Download - 2 2021 - Archiv-EuroMedica

ISSN 2193-3863

2 2021

ARCHIV EUROMEDICA ISSN 2193-3863

DisclaimerEuropäische Wissenschaftliche Gesellschaft e.V. HannoverSutelstr. 50A, 30659 Hannover, Deutschland

Tel. 49(0)5113908088Fax 49(0)511 3906454

Vorstand Dr. G. Tyminski, VorsitzenderEingetragen ins Vereinsregister am Amtsgericht Hannover: VR 7957

Design & layout by Tří barvy, s.r.o. Mariánské Lázně, Česká Republika

Editor-in-ChiefDr. Georg Tyminski EWG e.V., Hannover, Germany

Prof. Dr. Jörg Schulz Geriatric Clinics Berlin-Buch, Germany

Publishing DirectorProf. Aleksei Zhidovinov Astrakhan State Medical University, Astrakhan, Russia

Executive EditorProf. Dmitry Domenyuk Stavropol State Medical University, Stavropol, Russia

Managing EditorProf. Maya Dgebuadze Tbilisi State Medical University, Tbilisi, Georgia

Ethics ManagerProf. Gayane Khachatryan Erivan State Medical University, Erivan, Armenia

Associate EditorsProf. Sergey Kolbasnikov Tver State Medical Academy, Tver, Russia

Dr. rer. Nat. Stephan Heymann Noventalis – Institut für systemische BioKorrektur, Berlin-Buch, Germany

Editorial Advisory BoardProf. Nurlan AkhparovScientific Center of Pediatrics and Pediatric Surgery, Almaty, Kazakhstan

Prof. Vadim AstashovPeoples' Friendship University of Russia, Moscow, Russia

Prof. Tatiana BelousovaPrivolzhsky Research Medical University, Nizhny Novgorod, Russia

Prof. Eduard BatkaevPeoples' Friendship University of Russia, Moscow, Russia

Prof. Sergey DmitrienkoPyatigorsk Medical and Phamaceutical Institute, Pyatigorsk, Russia

Prof. Carlos Kusano Bucalen FerrariFederal University of Mato Grosso, Barra do Garças, Brazil

Prof. Iryna KhozhyloDnipropetrovsk Regional Institute for Public Administration Dnipro, Ukraine

Prof. Liana GogiashviliIvane Javakhishvili Tbilisi State University, Tbilisi, Georgia

Prof. Habibulo IbodovInstitute of Postgraduate Medical Studies, Dushanbe, Tajikistan

Prof. Vladimir IzranovImmanuel Kant Baltic Federal University, Kaliningrad, Russia

Prof. Igor V. KastyroMoscow S.U. Witte University, Moscow, Russia

Prof. Gulnara KapanovaAl Farabi Kazakh National University, Almaty, Kazakhstan

Prof. Semen KireevTula State Medical University, Tula, Russia

Prof. Vladimir KrestyashinPirogov Russian National Research Medical University, Moscow, Russia

Prof. Sergey LevakovI.M. Sechenov First Moscow State Medical University, Moscow, Russia

Prof. Nikogos OganesyanAcademy of Medical Sciences, Erivan, Armenia

Prof. Oral OspanovMedical University “Astana”, Astana, Kazakhstan

Prof. Ants PeetsaluTartu University Clinics, Tartu, Estonia

Prof. Urij PerestaUzhhorod National University, Uzhhorod, Ukraine

Dr. Olga PitirimovaMD, Bakulev Scientific Center of Cardiovascular Surgery, Moscow, Russia

Dr. Joerg PoetzschSatina Medical UG, Berlin, Germany

Prof. Vladimir ProtsenkoInstitute of Traumatology and Orthopedics NAMS of Ukraine, Kiev, Ukraine

Prof. Galina RevaBiomedicine School FEFU, Vladivostok, Russia

Prof. Stefan Antonio SanduStefan cel Mare University of Suceava, Iasi, Romania

Prof. Vaqif Bilas oğlu ShadlinskiyAzerbaijan Medical University, Baku, Azerbaijan

Prof. Natalia ShnayderKrasnoyarsk State Medical University, Krasnoyarsk, Russia

Prof. Rudolf Yuy Tsun-ShuKazakh National Medical University, Almaty, Kasachstan

Dr. Will Nelson VanceBeelitz-Heilstaetten Hospital for Neurological Rehabilitation, Beelitz, Germany

Dr. Mayank VatsRashid Hospital, Dubai Health Authority, Dubai, UAE

1| a r c h i v e u r o m e d i c a | 2 0 2 1 | v o l . 1 1 | n u m . 2 |

C O N T E N T S

EDITORIAL ........................................................................................... 4

P U B L I C H E A L T H

Pavel Seliverstov, Sofia Bakaeva,Daredzhan Tsurtsumiia, Valentin Shapovalov

TELEMEDICINE AS AN ESSENTIAL PARTOF THE MODERN HEALTHCAREDEVELOPMENT .................................................................................... 5

B I O C H E M I S T R Y

Oksana Boiko, Aleksandr Nikolaev,Daria Kozak, Natalia Gudinskaya,Maksim Sakharov, Polina GudinskayaYuri Dotsenko

ISOLATION AND PURIFICATION OF PROTEINTHAT IS IMMUNOLOGICALLY SIMILARTO HUMAN LACTOFERRIN ............................................................... 10

M O R P H O L O G Y , P H Y S I O L O G Y , P A T H O L O G Y

Artur Ovsepyan, Alexander Smirnov,Sergey Dydykin, Yuriy Vasil’ev,Evgeniy Trunin, Olga Shatunova,Aleksandr Aleksandrov, Angelina Ostyakova,Andrey Utkin

PERSONALIZED FSI-MODELING OF THE AORTIC BULBAND ARCH TO PREDICT ITS MECHANICALBEHAVIOR AND ASSESS THE LOADSDURING THE CARDIAC CYCLE ........................................................... 13

Oleg Zayko, Vadim Astashov,Anna Sindireva, Karina Basnakyan,Arina Lukyanchikova

EFFECT OF LEAD ON STRUCTURAL CHANGESIN LIVER OF WISTAR RATS UNDERTHE CONDITIONS OF ACUTE EXPERIMENT ...................................... 17

Anahit Karapetyan, Ashot Dallakyan,Marina Porchia, Carlo Santini,Gayane Khachatryan, Vahan Grigoryan

INFLUENCE OF COPPER COMPLEXES[CU(PTA)

4[BF

4] AND CU(II)

2(3,5-DIPS)

4(H

2O)

3

ON THE ORGANISM OF RATS IRRADIATEDWITH RADIOISOTOPE TECHNETIUM ................................................ 20

Gulnar Bayburina, Elena Nurgaleeva,Ajgul Samigullina, Ekaterina Farshatova,Eduard Agletdinov, Tatyana Tarasova,Larisa Mosina, Sergey Ryzhakin,Dalila Khaydar

EVALUATION OF THE CORTICOSTEROID RECEPTORS’ LEVEL IN THE KIDNEYS OF RATS AFTER SYSTEMIC ISCHEMIA-REPERFUSION ............................................... 23

Andrew Martusevich, Giulnar Orudzhova,Anastasia Romanova, Oksana Shubina

BACTERIAL SYMBIOSIS IN COMPLICATEDULCERS: THE PATHOGENETIC HYPOTHESIS ..................................... 26

P H A R M A C O L O G I C A L S T U D I E S

Anastasia Gerashchenko, Natalia Shabanova,Marina Efremova, Andrey Voronkov

STUDY OF POSSIBLE MECHANISMS OF ACTOPROTECTIVE ACTION OF СATEKHIN HYDRATE .................................................................... 28

Pavel Khoroshutin, Galina Reva,Tatsuo Yamamoto, Ivan Reva

PHARMACOKINETICS AND PHARMACODYNAMICS OF CHAGA BIRCH MUSHROOM COMPONENTS (INONOTUS OBLIQUUS) ........................................... 31

G Y N E C O L O G Y A N D O B S T E T R I C S

Yuliana Ukhanova, Adelya Zoeva,Olga Tishkova, Svetlana Belyavskaya,Adilya Ayupova, Pavel Loginov

BIOCHEMICAL STUDY OF MENSTRUAL BLOOD AND A MATHEMATICAL MODEL FOR DIAGNOSIS OF UTERINE MYOMAS ........................................... 39

S U R G E R Y

Oleksandr Burianov, Volodymyr Protsenko,Audai Abudayeh, Volodymyr Chornyi,Volodymyr Konovalenko, Yevhen Solonitsyn

THE RESULTS OF USING A BIOACTIVE GLASS-BASED COATING BY DEPOSITION ON THE CONTACT SURFACE OF PLATES IN BONE FRACTURES ASSOCIATED WITH TUMORS ............................................................. 43

2 | a r c h i v e u r o m e d i c a | 2 0 2 1 | v o l . 1 1 | n u m . 2 |

C O N T E N T S

Givi Odishelashvili, Sergei Shashin,Dmitri Pakhnov

RESULTS OF SURGICAL TREATMENT OF PATIENTS WITH PANCREATIC INJURIES ............................................................................................ 50

Aleksandr Gerasimov, Valeriy Nikolskiy,Aleksandr Mitroshin, Sergey Frolov,Ekaterina Titova

COMPLICATIONS OF OPEN SURGICAL PROCEDURES IN PATIENTS WITH PANCREATIC CYSTS ................................................................. 53

Vladimir Akimov, Dmitry Krikunov,Dmitry Parshin, Behruz Radzhabov,Vladimir Senko

LITERATURE REVIEWMETHODS FOR PROSTHETIC MESH FIXATION IN SURGICAL REPAIR OF INGUINAL HERNIAS ......................................................................... 56

Alexander Khitaryan, Alexander Murlychev,Alexey Orekhov, Sergey Kovalev,Vyacheslav Mykhaylichenko, Dmitry Parshin

EXPERIENCE OF ENDOVASCULAR EMBOLIZATION FOR RECTAL BLEEDING IN PATIENTS WITH RECTAL CANCER ...................................................................... 60

Denis Ushmarov, Sergey Gumenyuk,Alexander Gumenyuk, Tatyana Gaivoronskaya,Timofei Grigoriev, Ksenia Lukanina,Timofey Patsaev, Dmitry Domenyuk

PRECLINICAL TRIALS FOR ADVANCED CHITOSAN-BASED COATINGS IN TREATINGPURULENT WOUNDS......................................................................... 63

G A S T R O E N T E R O L O G Y / C L I N I C A L R E S E A R C H

Mikhail Turovets, Alexander Popov,Viktor Mandrikov, Yuriy Vedenin,Andrej Ekstrem

PREVENTION OF POST-ERCP PANCREATITIS IN HIGH-RISK PATIENTS ................................................................... 72

P E D I A T R I C S

Nikolay Cherkasov, Yuliya LutsenkoCLINICAL SIGNIFICANCE OF LAMININ AND ELASTINE LEVELS IN CHILDREN WITH UNDIFFERENTIATED CONNECTIVE TISSUE DISEASE ................................................................................ 78

Alexandra Popovicheva, Andrew Martusevich,Elvira Fedulova

FUNCTIONAL PROPERTIES OF PLATELETIN CHILDREN WITH IRRITABLE BOWEL DISEASE ................................................................................ 80

T H E R A P Y

Nikolay Sturov, Sergey Popov,Alina Kostenko, Grigory Roitberg

CHARACTERISTICS OF GUT MICROBIOTA IN AUTOIMMUNE DISEASES OF THE THYROID GLAND AND THE METHODS OF ITS CORRECTION .......................................................................... 82

Maria Mikhailova, Olga PoselyuginaBISMUT-INDUCED ONSET OF ACUTE TUBULOINTERSTICIAL NEPHRITIS: А СLINICAL CASE .............................................................................. 86

D E N T I S T R Y

Petr Petrov, Sergey Averyanov,Sergey Lazarev, Ilgiz Iskhakov, Adel Isaeva

EXPRESS METHOD FOR TREATING NEGATIVE EMOTIONAL STATES IN DENTISTS .................................. 89

Yuliya Makedonova, Dmitry Mihalchenko,Oksana Kurkina, Olga Afanasyeva,Sergey Veremeenko, Elena Venskel

IMMUNOLOGICAL ASSESSMENT OF THE STRESS RESPONSE IN PATIENTS WITH INFLAMMATORY POSTPROTHETIC COMPLICATIONS ............................................................................... 93

Dmitry Kompantsev, Anna ChahirovaRuslan Yusupov, Ilya Zaharchanko,Natalia Shabanova

DEVELOPMENT OF A BIODEGRADABLE MATERIAL FOR BONE REGENERATION BASED ON SILICON COMPOUNDS ................................................................ 98


Marina Suvorova, Galina Emelina,Anna Teplova, Belousova Anna

THE PREVALENCE OF NON-CARIOUS TOOTH DEFECTS ASSOCIATED WITH INDUSTRIAL EXPOSURE IN RESIDENTS OF PENZA REGION (RUSSIA) ................................. 101

Vladimir Tlustenko, Valentina Tlustenko,Sergey Komlev, Alexander Ivaschenko,Vladimir Koshelev

IMPLANT TREATMENT IN PATIENTS WITH SEVERE TOOTH WEAR ON THE OCCLUSAL SURFACE .......................................................... 104

Natalya Prozorova, Roman Fadeev,Victor Weber, Marina Chibisova,Natalia Robakidze, Irina Prozorova,Sofya Sokolova, Vladimir Shkarin

EVALUATING OPTICAL DENSITY OF ALVEOLAR BONE IN PATIENTS WITH DIABETES MELLITUS USING CONE-BEAM COMPUTED TOMOGRAPHY ................................................................................ 108

Marietta Sukhorukikh, Marina Kozlova,Ekaterina Gorbatova, Larisa Dzikovitskaya,Aleksey Bashtovoy

CLINICAL COURSE OF PERIODONTAL DISEASES IN PATIENTS WITH POSTMENOPAUSAL OSTEOPOROSIS, TREATED WITH BISPHOSPHONATES .............................................................. 118

Artem Kashtanov, Hadi Darawsheh,Yuriy Vasil’ev, Sameh Ibrahim,Darina Shirokova, Ekaterina Shilyaeva,Valeria Brichagina, Guseyn Taymurazov

INFERIOR ALVEOLAR NERVE BLOCK BY INJECTION INTO THE PTERYGOMANDIBULAR SPACE USING GUIDING DEVICES: A SYSTEMATIC REVIEW ........................................................................................... 123

Yuliya Makedonova, Alexander Vorobyev,Anna Osyko, Alexey Mikhalchenko,Alexander Alexandrov, Anastasia Pavlova-Adamovich

CLINICAL AND ANATOMICAL REQUIREMENTS AND INDICATIONS FOR USING A NOVEL MULTIFUNCTIONAL GNATIC DEVICE ............................................................................... 130

https://orcid.org/0000-0001-8223-2697


E D I T O R I A L

Dear clinicians, researchers, colleagues and friends!

Treatment of wounds poses acute medical and social challenges for research and practice. It acquires even more importance with an increase in natural dis-asters, industrial and domestic accidents and military conflicts.

In modern medicine there are a great number of wound dressings. Their application is based on the principle of moist recovery of the wound formulated by G. Winter (1962) and C.D. Hinman, Н. Maibach (1963). The authors established that moist antibacte-rial environment was optimal for maintenance of nor-mal mitotic process in the wound: migration of epider-mal cells and epithelialization of wounds occur more actively in the moist environment and not in the open air which dries out the surface. The moist environment ensures a higher protease activity that enables prompt cleaning of wound surface without use of fermenta-tion drugs associated with allergic response. Besides, it prevents dryness of nerve endings and relieves pain. As the dressing does not stick to the wound, there is no pain on removal and no damage to granulation tissue and growing epithelia.

A dominant tendency in the development of modern wound dressings is the use of polymer matrix composites based on biocompatible natural and synthetic polymers. Effective healing of wounds and wound infections requires dressings to have a high sorption activity, effectively removing excessive wound exudate and its toxic components. Such dressings are semi- permeable to vapor and at the same time prevent dryness of the wound surface and protect from

Dmitry Domenyuk

Executive Editor,Archiv EuroMedica

Wound Management with Chitosan Based

Biomaterials

secondary infection of the wound; they are elastic and capable of restoring surfaces with complicated relief, and they are mechanically robust. Dressings should provide an optimal microenvironment for wound recovery and cause neither pyrogen and toxic effects nor irritant and allergic reactions.

One of the advantages of dressings based on composite matrices is their ability to deliver antibacte-rial, antiseptic, anti-inflammatory drugs that facilitate reparative processes. Transparent films allow visualiza-tion of the wound bed.

Absorbable membranes made from polymer matrices mostly conform to all clinical specifications and could be applied at all stages of wound and burn treatment. Optimal mechanical and physicochemi-cal properties of hydrogel compositions used in drug delivery enable a prolonged effect of drug substances, targeted drug delivery and possibly a marked synergis-tic effect of drugs.

Chitosan and its derivatives are viewed as most promising biopolymers. Chitosan is not toxic, it is biocompatible and biodegradable. It has antimicrobial and immunostimulating activities. Chitosan is a he-mostatic agent, it accelerates coagulation process, and blocks nerve endings alleviating the pain. Chitosan has been actively investigated as the basis of wound and burn dressings and a major component of multiple products: hydrogels, membranes, films, nanofibers, granules, nanoparticles and scaffolds. Scaffolds are of particular interest as new universal forms which can be utilized in tissue engineering and regenerative medi-cine.

In the section Surgery you will be offered the data on effectiveness of novel developments in chitosan-based dressings used for treatment of biliary and heavy exudate wounds. Results of researches broaden our knowledge on application of chitosan-based wound dressings and their anti-inflammation and regenerative properties.

http://dx.doi.org/10.35630/2199-885X/2021/11/2ed

http://dx.doi.org/10.35630/2199-885X/2021/11/2ed


TELEMEDICINE AS AN ESSENTIAL PART OF THE MODERN HEALTHCARE DEVELOPMENT

A B S T R A C T — Widespread use of telemedicine as a method for remote medical care provision can become one of the most promising ways to optimise modern healthcare. This article proposes a methodology for adult health telemedicine screening using a system for analysing unstructured data of monitoring chronic non-communicable diseases in the population. This method is based on the risk estimation by the Decision Rules approach, and the fuzzy set theory is used as a description tool. Scientifically-based comprehensive clinical approach for the development of medical questionnaires, the knowledge of which depends on a large scientific base in the field of screening, propaedeutic, taking into account the standards and requirements of health assessment in the Russian Federation, is a fundamental difference from the existing analogues. Expert system of telemedicine questionnaire screening allows determining the health risks by specific profiles, to develop a final personalised judgment with recommendations for a healthy lifestyle, further examination, treatment, and prevention of chronic non-communicable diseases.

K E Y W O R D S — telemedicine, screening, chronic diseases, health protection, quality improvement in healthcare, information and communication technologies (ICT).

Pavel Seliverstov1 , Sofia Bakaeva1 ,Daredzhan Tsurtsumiia1 , Valentin Shapovalov2

1 I.I. Mechnikov North-Western State Medical University, Saint Petersburg; 2 Saint Petersburg State Electrotechnical University “LETI”, Saint Petersburg, Russia

[email protected]

B A C K G R O U N DCurrently, there is a steady increase in public

spending related to healthcare worldwide. Widespread introduction of telemedicine in general practice as a method for provision of qualified remote medical care is one of the promising options for optimising such costs.

Telemedicine has existed for a long time one way or another. E.g., in 1905, Einthoven first transmitted ECG records using telephone communication. Since 1922, the University Hospital in Gothenburg has been providing medical consultations to seafarers via radio channels. In 1959, the first television consultation of a psychiatric patient was held in the United States. Later

Received 04 March 2021 Received in revised form 15 April 2021; Accepted 25 April 2021

in 1965, the American cardiac surgeon, M. Debeki, consulted surgeons, performing a surgery in Geneva, via a satellite communication channel. In 1999, Ms Gerry Nielsen, engaged as a medical specialist at the Amundsen-Scott Research Station in Antarctica, was diagnosed with and treated for breast cancer through satellite communication, email, and video conferencing. [1].

According to WHO, the term telemedicine defines health services provision in conditions where the distance is a critical factor by healthcare professionals, using information and communication technolo-gies (ICTs) to exchange the necessary data for the diagnosis, treatment and prevention of diseases and injuries, conduct research and assessments, as well as to continue the education of health professionals for the improvement of public health and the develop-ment of local communities [2]. In turn, the European Commission considers this term more specifically and proposes to construe telemedicine as operational remote access to the services of medical specialists, us-ing ICTs, regardless of the patient location or the way the relevant information is stored. [EHTEL, 2008]. According to the definition of the American Tele-medicine Association (ATA), telemedicine is the use of medical information provided by one party to another by electronic means of communication to improve the health of patients. [3] Despite the fact that each insti-tution offers its own interpretation of the term, each of the definitions highlights three main characteristics of telemedicine: health care quality improvement, the use of ICT, and remote access. [4].

In 2010, WHO identified four specific principles of telemedicine [WHO 2010]: provision of clinical support, overcoming geographical barriers and estab-lishing communication between physically distant users, using various ICTs, improvement of public health [2].

S Y N C H R O N O U S A N D A S Y N C H R O N O U S T E L E M E D I C I N E

Telemedicine, depending on the terms of in-formation transmission and interaction between the persons involved in the process, can be divided into 2 types: synchronous and asynchronous [2]. Asynchro-


http://dx.doi.org/10.35630/2199-885X/2021/11/2/1

http://orcid.org/0000-0001-5623-4226

http://orcid.org/0000-0002-4297-8918

https://orcid.org/0000-0001-7806-9364

https://orcid.org/0000-0002-9764-4018

http://dx.doi.org/10.35630/2199-885X/2021/11/2/1


nous telemedicine is based on the exchange of pre-re-corded data between two or more persons at different periods of time. For example, the sending a clinical case to an expert via email with subsequent doctor's feedback on the diagnosis, optimal therapy, and rec-ommendations for a healthy lifestyle management can serve as the examples of asynchronous telemedicine, Synchronous telemedicine is applicable in real time and requires simultaneous presence of participants in the process, e.g., during video conferences, phone conversations or via the Internet. In both synchronous and asynchronous telemedicine, information can be transmitted in various forms: as text, audio, video, or image [5].

R E M O T E M O N I T O R I N G A N D C O V I D - 1 9

Remote monitoring can be singled out separately. This is a form of telemedicine that involves remote monitoring of the patient's condition by medical professionals using various technical devices. The data obtained during monitoring is subject to remote monitoring and is transmitted to a medical facility for further analysis and interpretation. [4, 6] Remote monitoring is used for dynamic observation of pa-tients with cardiovascular diseases, diabetes mellitus, bronchial asthma, as well as for domiciliary patients monitoring. [3]

The use of modern technologies now makes it possible to provide high-quality medical care without violating the principles of social distancing, which is really important in the context of the COVID-19 pan-demic, where telemedicine has become the first line of defence for both patients and medical specialists. [7] As practice has shown, telemedicine can be used in several forms in conditions of global quarantine [7]:

1. Online consultations: tele- and video confer-ences;

2. Telemonitoring: the use of screening devices, which assess the level of blood pressure, satura-tion as well as the respiratory rate;

3. Chatbots, where the patient can get answers to frequently asked questions, as well as request con-nection with the doctor.

Nowadays, telemedicine combines convenience, low cost, and high availability for the end user. As a result, it reduces the time for diagnosis and treatment, including in the context of the COVID-19 pandemic, as well as possibility of close observation without ne-cessity of moving around the city, and direct physical contact with a medical professional, which ultimately reduces the risk of spreading the infection. In addition, telemedicine can be used to control the spread of both

non-communicable and infectious diseases, including COVID-19 [8, 9, 10].

Recent events related to the COVID-19 pan-demic, forced self-isolation regime, cancellation of face-to-face appointments and re-profiling hospitals have given impulse for development of telemedicine in Russia. The population began to show more and more interest in telemedicine, considering that after the be-ginning of COVID-19 the need for medical consulta-tions did not disappear, but, on the contrary, increased. Thus, in 41% of cases, a face-to-face consultation was not required after a remote request for medical care, 20% of those who applied were re-admitted to the same specialist and in 30% of cases, and patients were immediately referred to a narrow-profile specialist without another consultation with a therapist [11].

T E L E M E D I C I N E M A R K E T A N D S E R V I C E S

Currently, the development of telemedicine services is an absolute trend in the development of the digital industry worldwide. According to various estimates, the average annual growth rate of recently emerging online medical opportunities for users in the next five years will be about 116%. If the volume of the Russian telemedicine market was estimated at RUB 1.5 bln in 2019, it is expected to grow more than 60 times by 2025, reaching RUB 96 bln, which is consist-ent with the VEB Ventures opinion. At the same time, the company notes that the growth driver will be not just the service demand dynamics effect, but also the availability of telemedicine platform services for a wide range of patients [12, 14].

In terms of the global market, according to BBC Research, the global telemedicine market increased to an average of $44 bln in 2019. According to P&S Mar-ket Research in 2016, its volume did not exceed $18 bln. Lower market activity in developing countries is related to insufficient payback and lack of technologi-cal infrastructure. In developed countries, telemedicine complements traditional medicine, while in develop-ing countries telemedicine is often an alternative to traditional medical care. Despite this fact, the emer-gence of telemedicine in developing countries opens up access to health services for a wide range of people, as well as reduces geographical barriers [12].

We analysed the market of companies, developing telemedicine, and found 35 services, and almost all of them interact with the client by means of a video cam-era (video service), smartphone, or computer [13–20]. The price of such services in the foreign market ranges from $5 per month to $441 per insurance package per year. For comparison, in 2020, the average check in private clinics in the Russian Federation was amounted



to RUB 3,000 (€33) per visit, and the average cost of a minimum check-up: therapist's appointment, blood and urine tests, lung X-rays and ECG was amounted to RUB 40,000 (€442). The price of a specialist consulta-tion using telemedicine technologies on the Yandex-Health portal starts from RUB 499 (€5.5). The main type of services provided is the patient flow control, remote telemedicine counselling, and signal processing from gadgets [21].

Thus, in terms of cost-effectiveness, it is obvi-ous that the introduction of telemedicine will reduce health care costs [22, 23]. The reduction in health care costs will be achieved by reducing the cost of hospi-talisation, patient transportation, and indirect costs associated with leaving home or office [24]. Thus, a number of Russian and foreign studies have shown a reduction in costs for telemedicine in comparison with traditional methods from 2 to 73%. Even now, telemedicine consultations for the end user have be-come over 10 times more profitable than face-to-face consultations [22, 25].

Thus, there are all the prerequisites for the emer-gence of own developments using telemedicine tech-nologies in Russia. However, among the well-known programs, along with their undoubted advantages, such as accessibility to the public, possibility of remote examination, relatively low cost of screening, there are also disadvantages such as lack of final analytical docu-ments, coverage of one or more systems, need for a face-to-face visit to the doctor to estimate the risks and determine further tactics of patient control [26].

M A T E R I A L S A N D M E T H O D SWe have developed our own version of telemedi-

cine questionnaire screening for adult health, based on the experience of national and foreign colleagues. The scientific-based comprehensive clinical approach to the development of the medical questionnaire, the knowledge of which is based on a large scientific base in the field of screening, propaedeutic, taking into account the standards and requirements of the health assessment of the Russian Federation, is a fundamental difference from the existing analogues. Creation of a decision tree model and development of algorithms for risk share estimation, taking into account external factors, including the specifics of the residence halo, cultural characteristics of nutrition, and others, will improve the quality of medical screening, its reli-ability, and will also make it possible to give the user optimal recommendations on the strategy of diagno-sis and treatment. Over the past year and a half, the authors have done a great deal to create a telemedicine system of questionnaire screening, a methodology for selection and ranking of questions that are most in-

formative, simple and available for stating, taking into account an integrated approach [27, 28].

The study was conducted at the Department of Internal Medicine, Clinical Pharmacology and Neph-rology of the North-Western State Medical University named after I.I. Mechnikov, based on our platform for telemedicine medical support. The study involved 139 people: 97 women and 42 men, the average age of the subjects was 65±13.8 years.

We did not use a clinical entity as a health risk assessment in the telemedicine system, but rather a pathology profile, in which a set of analytical features, objective examination data, and complaints were divided into groups. 198 questions were prepared for conducting a multidisciplinary comprehensive questionnaire health screening; these questions were structured according to 5 pathology profiles: cardiol-ogy, endocrinology, gastroenterology, pulmonology, and oncology. These pathology profiles were not chosen by chance: Since the second half of the 20th century, the main death causes have been chronic non-communicable diseases (chronic NCDs): circulatory illnesses, chronic bronchopulmonary diseases, onco-logical diseases, and diabetes mellitus. In the Russian Federation, about 75% of all deaths occur annually due to chronic non-communicable diseases, however, particularly high mortality is observed among people of working age [29]. In addition, the prevalence of digestive diseases has increased in recent years [30].

R E S U L T S A N D D I S C U S S I O NAll the fairly ascertained features of each profile

were evaluated according to the degree of their severity or the reliability of their presence. Minimal deviations were taken for early detection of pathology or low risk, in cases of its combination with any complaints or de-velopmental features noted in the questionnaire. The program provides both identification of the severity degree and attribution of the symptom or sign to the pathology of several systems, since they can often appear during destruction of different organs, and therefore the estimation of the disease risk spectrum is carried out by the system in accordance with all five profiles. In turn, the system is configured for the boundary between the risk group and the pathology, estimated at 50 points, and the range of the risk group is concentrated in the range from 20 to 50 points. The risk estimation is carried out by the Decision Rules method, and the fuzzy set theory is used as descrip-tion. The typical features of the mathematical appara-tus used are, on the one hand, the ability to formalise ideas about the degree of severity of a particular symp-tom, and on the other hand, the adequacy of medical logic.



Using the developed technology, it was found that 33.8% of the subjects were diagnosed with a low risk of cardiovascular diseases, 41.7% — an average risk, 24.5% — a high risk. While analysing the answers to the questions of the gastroenterological profile, 33.1% had a low risk of gastrointestinal diseases, 46.8% had an average risk, and 20.1% had a high risk. 46.7% of the subjects had a low risk according to the Pulmonology profile, 40.3% had an average risk, and 13% had a high risk. 6.5% of the respondents were diagnosed with low risk in the Endocrinology profile, 23.7% were diagnosed with medium risk, and 69.8% were diagnosed with high risk, however, this profile re-quires further development due to the non-specificity of endocrinological complaints. 18% of the subjects have a low risk of cancer, 32.4% have an average risk, and 49.6% have a high risk of cancer, according to the questionnaires, which requires increased cancer alertness in this study group. The average score of the subjects in the cardiological profile was 517, in the gas-troenterological profile — 440, the average value in the Pulmonology profile was 439, the average value in the Endocrinology profile was 495, and the average value in the Oncology profile was 533. 12 subjects (8.6%) were revealed to have a high risk in all 5 pathology profiles. And only 7 (5%) of the examined patients were diag-nosed with a low risk for all pathology profiles.

The program analysed the obtained data and offered appropriate recommendations depending on the age, gender, body mass index, vicious habits, and level of physical activity and identified risk factors of the subject. Thus, in case of a low risk of disease development in all 5 pathology profiles, it is recom-mended to follow healthy lifestyle rules, including the principles of proper nutrition, body weight control, sufficient physical activity, rejection of vicious habits and medical examinations according to the established deadlines. In case of an average risk of disease develop-ment, depending on the pathology profile, in addition to the healthy lifestyle, it is recommended to monitor blood pressure daily, as well as to determine a set of necessary laboratory and instrumental examinations, and immunisation. In case of high risk of pathologies, the program recommends a full-time consultation with a doctor to determine the further strategy of examination and treatment of such a patient, as well as a dispensary follow-up with an appropriate specialist at the place of residence.

C O N C L U S I O NThus, our program makes it possible to determine

the degree of existing health risk by specific profiles (cardiology, gastroenterology, pulmonology, endo-crinology, oncology), to develop a final judgment

and recommendations for a healthy lifestyle, further examination, treatment and prevention of chronic diseases.

The innovation of this development lies in in-house methodology for the selection and ranking of clinical symptoms and signs of diseases, taking into account their severity and reliability. The specified symptoms and signs of diseases create not a clinical entity, but a pathology profile, and therefore they have the greatest information content, statement availabil-ity, and cover all the key body systems. Moreover, an original case of summary documents with recommen-dations on lifestyle changes has been developed.

The benefits of this development are the flexibil-ity of the decision rules, corresponding to the opinion of the expert doctor, the convenience and clarity of the representation of the final results, the formula-tion of conclusions on the necessary follow-up and recommendations for adherence to a healthy lifestyle, as well as individualised approach, mobility, absence of reference to time and place, and the availability of doctor–patient feedback.

AcknowledgmentProject for the development of an intellectual

telemedicine platform became the winner of the UM-NIK (a smart person) competition within the frame-work of the Digital Economy of the Russian Federation national program.

FundingThis study is supported by the Innovation Assist-

ance Fund under Agreement No. No. 66GUTSES8-D3/56432 dated 21.12.2019.

R E F E R E N C E S1. Alsup D. Doctor rescued from Antarctica in 1999

dies at 57 [Electronic] CNN2009 – URL: http:// www.cnn.com/2009/HEALTH/06/23/obit.jerri.nielsen/index.html (reference date: 01.03.2020).

2. World Health Organization. Report on the Second Global Survey on eHealth. 2012. ISBN 978-92-4-456414-1.

3. American Telemedicine Association: official website. – Washington. – URL: https://www.americantelemed.org (reference date 01.03.2020).

4. Kim J., Alanazi H., Daim Т. (2015) Prospects for Telemedicine Adoption: Prognostic Modeling as Exemplified by Rural Areas of USA. Foresight and STI Governance, vol. 9, no 4, pp. 32–41. DOI: 10.17323/1995-459x.2015.4.32.41.

5. Rao B, Lombardi A II. Telemedicine: current status in developed and developing countries. Journal of Drugs in Dermatology, 2009, 8(4):371–375. URL: http://docs.cntd.ru/document/1200157018 (refer-ence date 09.04.2020).



6. Weinstein RS, Krupinski EA, Doarn CR. Clinical examination component of telemedicine, telehealth, mhealth, and connected health medical practices. Med Clin North Am 2018 May;102(3): 533–544. doi: 10.1016/j.mcna.2018.01.002.

7. Vidal-Alaball J, et al. Telemedicine in the face of the COVID-19 pandemic. AtenPrimaria. 2020 https://doi.org/10.1016/j.aprim.2020.04.003.

8. Xia W et al. Transmission of coronavirus disease 2019 during the incubation period may lead to a quar-antine loophole. medRxiv (preprint 2020) https://doi.org/10.1101/2020.03.06.20031955.

9. Hollander JE, Carr BG. Virtually Perfect? Telemedicine for Covid-19. New England Journal Of Medicine. 2020; https://www.nejm.org/doi/full/10.1056/NEJMp2003539 .

10. Lurie N, Carr BG. The Role of Telehealth in the Medical Response to Disasters. JAMA Inter-nal Medicine. 2018;178(6):745. https://jaman-etwork.com/journals/jamainternalmedicine/article-abstract/2678828. doi: 10.1001 / ja-mainternmed.2018.1314

11. Kiseleva A. Just to Ask: Coronavirus Has Boosted The Demand For Online Doctors In Russia. Analytical articles. 2020. Access mode: https://1prime.ru/state_regulation/20200519/831468461.html (reference date 21.02.2021).

12. Combi, Carlo et al. “Telemedicine for Developing Countries. A Survey and Some Design Issues.” Applied clinical informaticsvol. 7,4 1025–1050. 2 Nov. 2016, doi:10.4338/ACI-2016-06-R-0089.

13. Oscar – Taking on Giants – Health Insurance 2.0: official website. – URL: https://digital.hbs.edu/platform-rctom/submission/oscar-taking-on-giants-health-insurance-2-0/ (reference date 26.02.2021).

14. ODYSSEY Clinical Decision Support: official web-site. – URL: https://www.oneadvanced.com/solu-tions/odyssey/ (reference date 26.02.2021).

15. Google.health: official website. – URL: https://health.google (reference date 26.02.2021).

16. Medwhat: official website. – URL: https://medwhat.com (reference date 26.02.2021).

17. SberZdorovie: official website. – URL: https://spb.docdoc.ru (reference date 26.02.2021).

18. Helfine Medical: official website. – URL: https://www.helfine.ru (reference date 26.02.2021).

19. Health.mail: official website. – URL: https://health.mail.ru (reference date 26.02.2021).

20. Yandex.Health: official website. – URL: https://health.yandex.ru (reference date 26.02.2021).

21. Manukian E. How The Pandemic Changed Commercial Medicine? E. Manukian. Analytical articles. Rossiyskaya gazeta. 2021; Access mode: https://rg.ru/2021/02/14/kak-pandemiia-izmenila-kommercheskuiu-medicinu.html (reference date 21.02.2021).

22. Bongiovanni-Delarozière I., Le Goff-Pron-ost M. 2017. Economic evaluation methods applied to telemedicine: From a literature review to a standard-ized framework. European Research in Telemedicine. November 2017, 6(3–4): 117–135.

23. Isabel de la Torre-Díez, Miguel López-Coro-nado, Cesar Vaca, Jesús Saez Aguado, and Carlos de Castro. Telemedicine and e-Health.Feb 2015.81–85. http://doi.org/10.1089/tmj.2014.0053.

24. Dowie R., Mistry H., Rigby M., Young T.A., Weather-burn G., Rowlinson G. et al. 2009. A pediatric telecardiology service for district hospitals in south-east England: an observational study. Arch Dis Child. 94(4): 273–277. doi: 10.1136/adc.2008.138495. Epub 2008 Sep 11.

25. Levanov V. M., The Scientific Rationale of the Elec-tronic Technologies Using under the Public Health Modernisation Conditions at the Regional Level. Author’s abstract Diss. …M. D. 14.02.03. Moscow.

26. Baranov A. A., Vishneva E. A., Namazova-Baranova L. S. Telemedicine – Prospects And Difficulties Before a New Development Stage. Pedi-atricheskaya farmakologiya. 2013;10(3):6–11. https://doi.org/10.15690/pf.v10i3.691.

27. Seliverstov P. V., Bakaeva S. R., Shapovalov V. V. A Telemedicine System in the Assessment of Risks For Socially Significant Chronic Non-commu-nicable Diseases. Vrach. 2020; 31 (10): 68–73. https://doi.org/10.29296/25877305-2020-10-13.

28. Seliverstov P. V., Bakaeva S. R., Shapovalov V. V. Development of a Medical Methodology for Telemedicine Screening Adult Health in Outpa-tient Settings for Population Monitoring Chronic Non-communicable Diseases. Meditsynsky Sovet. 2020;(11):202–209. https://doi.org/10.21518/2079-701X-2020-11-202-209.

29. Boytsov, S. A. Prevention of Chronic Non-commu-nicable Diseases. Recommendations / S. A. Boitsov, A. G. Chuchalin, G. P. Arutyunov, et al. – M., 2013. – P. 5–6.

30. Denisova T.P., Shuldyakov V.A., Tyultyaeva L. A, Chernenkov Yu. V., Alipova L.N, Sadjaya L.A. Monitoring Internal Diseases Dissemination (Digestive Disorders) // Saratov Journal of Medical Scientific Research 2011. Vol. 7, No 4. P. 772–776.



ISOLATION AND PURIFICATION OF PROTEIN THAT IS IMMUNOLOGICALLY SIMILAR TO HUMAN LACTOFERRIN

A B S T R A C T — The study aimed at isolating a substance that is immunologically similar to human lactoferrin, hereinafter — microbial-derived lactoferrin (MdLF) isolated from K.pneumoniae liquid culture.Protein extraction started with isolation of ballast proteins in 2M ammonium sulphate (p.a.). Ion chromatography with cation-exchange agents was the basic method used for isolating MLF. Proteins from the isolated protein fractions were identified with AGID method. The isolated MLF is glucoprotein with the molecular mass of about 84,000, the agarose diffusion coefficient of 3.1 × 10–7 cm2 sec–1 and the relative electrophoretic mobility of 0.42. MLF is characterized by low hydrophobicity and elutes from phenyl-Sepharose with 0.4 ammonium sulphate. Its isoelectric point equals to 9.2.

K E Y W O R D S — microbial-derived lactoferrin, chromatography, electrophoresis.

Oksana Boiko1 , Aleksandr Nikolaev1 , Daria Kozak1 , Natalia Gudinskaya1 ,Maksim Sakharov2 , Polina Gudinskaya1 Yuri Dotsenko

1 Astrakhan State Medical University, Astrakhan;2 N.I. Pirogov City Polyclinic No 8, Astrakhan region, Russia

[email protected]

R E L E V A N C ELactoferrin belongs to the transferrin family and

is capable of active binding and transporting of Fе3+ and other mixed-valent metals. Previously examined lacroferrins are glycoproteins with the molecular masses within the range of 75–80 kDa, containing intramolecular disulfide bonds. A polypeptide chain of lactoferrin forms two globular domains called M- and C-lobes, connected with an alpha helix. Each lobe has an iron-binding centre. The tertiary structures of apo-lactoferrin (iron-free lactoferrin) and iron-binding lactoferrin differ [4, 6, 7, 8, 9, 10, 13]. For example, it is found that Fе3+ binding plays an essential role in implementing the antioxidant and bacteriostatic func-tion of lactoferrin. Along with antimicrobial activity, lactoferrin also has other biological properties — it can act as an immunomodulatory agent, antioxidant and anti-inflammatory agent and transcriptional

Received 22 April 2021; Received in revised form 26 May 2021; Accepted 29 May 2021

factor and definitely participates in iron metabolism [1, 5, 14, 15, 17].

Comparative studies play a significant role in examining biological properties of lactoferrin, for which reason it is important to examine the structure and properties of lactoferrin from the sources covering different species. Interestingly, literature sources still contain only sporadic data on the presence of lactof-errin in prokaryotes with no protein isolated and no molecular mass measured [2, 3, 10, 11, 12].

Anyway, microbial cells have been increasingly used as potent protein factories because of their versa-tility and cost-effectiveness [18].

We have made an attempt to isolate and purify a protein that is immunologically similar to human lactoferrin from Klebsiella pneumoniae. MdLF pres-ence in the samples of the pre-centrifuged (5,000 rpm during 45 minutes) liquid culture of K.pneumoniae (meat-peptone broth, incubation time — 24 hours at t=37° C) was assumed on the basis of EIA results with the use of Hycult biotech test kits (Netherlands) [3].

M A T E R I A L S A N D M E T H O D S Currently, lactoferrin is commonly isolated by the

method of ion chromatography with cation-exchange agents. This method is based on a significant difference of the LF isoelectric point from other proteins at the pH values of 7–7.4.

Protein extraction started from isolation of bal-last proteins in 2M ammonium sulphate (p.a.). MLF does not precipitate at this concentration, and once a precipitate was centrifugally isolated (10,000 rpm during 45 minutes), it was resuspended in a buffer (10 mM tris-НСl, pH 7.6; 1 mM EDTA; 1 mM iron(III)-ammonium) to saturate LFP with iron.

After this, the sample is thoroughly desalted and buffered with a phosphate buffer, рН=7.0. Then the obtained solution was applied on a column (4.5×60 cm) of Heparin Sepharose (Sigma, USA) which had been preliminarily equilibrated with 20 mM tris-НСl, pH 7.5. The sorbent was washed with 20 mM tris-НСl, pH 7.5, and then — with the same buffer, containing 1% triton X-100, and then again – with the buffer with no triton until the optical absorp-tion disappeared. Elution was carried out with NaCl linear gradient with the concentrations from 0 to 1 M


http://dx.doi.org/10.35630/2199-885X/2021/11/2/2

http://orcid.org/0000-0003-3882-9300

http://orcid.org/0000-0001-6607-430X

http://orcid.org/0000-0002-6653-7658

https://orcid.org/0000-0002-2643-4806

https://orcid.org/0000-0001-5135-9679

https://orcid.org/0000-0002-4989-491X

https://orcid.org/0000-0002-6016-7246

http://dx.doi.org/10.35630/2199-885X/2021/11/2/2


in 20 mM tris-НСl, pH 7.5. The obtained fractions were dialyzed against 10 mM tris-НСl, pH 7.5 at 4° С during 16 hours.

Unlike the general method of isolating MdLF, the heparin sepharose column had been preliminary treated with 0.2–0.3% formaldehyde solution in order to oxidize minor functional groups of heparin, which leads to an increase in its negative charge and, as a result, increases it affinity in relation to lactoferrin.

The molecular mass of lactoferrin was measured with DС-Na-polyacrylamide gel electrophoresis ac-cording to Schägger and von Jagow’s method with the use of 8% acrylamide gel. The molecular mass was also evaluated upon gel filtration results.

R E S U L T S The isolated MdLF is glucoprotein with the

molecular mass of about 84,000, the agarose diffusion coefficient of 3.1 × 10–7 cm2 sec–1 and the relative elec-trophoretic mobility of 0.42. LFP is characterized by low hydrophobicity and elutes from phenyl-Sepharose with 0.4 ammonium sulphate. Its isoelectric point equals to 9.2.

Despite its immunochemical identity with hu-man milk lactoferrin, MdLF differs from lactoferrin by some of its physiochemical parameters. The difference is especially evident in the electrophoretic mobility of MdLF and human milk lactoferrin. According to the obtained data, the MdLF relative electrophoretic mobility is 0.41±0.006, while the one of human milk lactoferrin is 0.47±0.003. These differences may be considered another evidence of the presence of several isoforms of lactoferrin that have the same molecular mass and are immunochemically similar but differ in their affinity to different ionogenic groups.

Dissociation of the LF and Fe3+complex was ex-amined with two methods — at low concentrations of chelating anions (Ре^ dissociation synchronically hap-pens from two iron-binding centres of LF) and at high concentrations of chelating anions (Fe3+ dissociation from two iron-binding centres happens separately). Iron-saturated LF (chololactoferrin) has a typical vis-ible absorption spectrum. The spectra appeared to be similar for the bacteria and human proteins. No differ-ence in the bacteria and human proteins was identified in the experiments on the LF and Fe3+ complex disso-ciation. Summary data on the MdLF physicochemical properties are presented in Table 1.

C O N C L U S I O N Modern research allows to state that iron is a uni-

versal factor of microbial growth. Being in the environ-ment, iron is exposed to oxidation and hydrolysis proc-esses, leading to a decrease in the concentration of Fe2+

Table 1. MdLF physicochemical properties

and Fe3+ free ions down to 10–9–10–18 M, which is not sufficient for optimal vital activity of most of the mi-croorganisms. Likewise, iron is bound to iron-binding proteins in the mammal organisms and is unavailable for microorganisms. That is why microorganisms have evolved ways of getting iron in iron-deficiency condi-tions, which is important for optimal development in environment and, in certain cases, may be critical for survival of the population. In this case, microbial strategies for getting iron, including MLF synthesis, can be considered as one of the factors that determine the formation and stability of microbial associations, playing an important role in the functioning of normal and pathological microbiocenoses in the host.

R E F E R E N C E S 1. Boiko O.V., Salko V.N., Boiko A.V., Martova

O.V., Akhmerova R.R., Sribny I.V., Tishina I. Yu. Effect of toxic substances on opportunistic patho-gens. Hygiene and Sanitation. 2002, No.1. P. 54–56.

2. Boiko O.V., Terentiev A.A., Boiko V.I. Molecu-lar mechanisms of bacteria carrying (characteristics and detailed analysis)/ Saarbrucken, 2012.

3. Boiko O.V., Dotsenko Yu.I., Gudinskaya N.I., Boiko V.I., Mukhamedzyanova R.I., Kozak D.M., Stenkin F.S. Effect of pollutants on bio-chemical properties of microorganisms. Hygiene and Sanitation. 2020; 99 (4): 368–378. doi: https://doi.org/10.33029/0016-9900-2020-99-4-368-378.

4. Brock JH. Lactoferrin – 50 years on. Biochem Cell Biol. 2012 Jun;90(3):245–51. doi: 10.1139/o2012-018.

5. Drago-Serrano ME, Campos-Rodríguez R, Carrero JC, de la Garza M. Lactoferrin: Balanc-

Properties LFPMolecular mass 85,000±5,000Relative electrophoretic mobility in agar 0.42±0.005

Agarose diffusion coefficient, cm2 sec -1 3.1 × 10 –7

Hydrophobicity Elution from phenyl-Sepharose with 0.4 ammonium sulphate

Precipitation with ammonium sulphate, % saturation and precipitation range

50–75

Full precipitation 75Precipitation with 50% ethanol No precipitationPrecipitation with 60% acetone Full (reversible)Precipitation with 0.9M chloric acid Irreversible

Isoelectric point 9.2Thermal resistance Resistant at 65° С — 30 minutes



ing ups and downs of inflammation due to microbial infections. Int J Mol Sci. 2017 Mar 1;18(3):501. doi: 10.3390/ijms18030501.

6. González-Chávez SA, Arévalo-Gallegos S, Rascón-Cruz Q. Lactoferrin: structure, func-tion and applications. Int J Antimicrob Agents. 2009 Apr;33(4):301.e1–8. doi: 10.1016/j.ijantimi-cag.2008.07.020

7. Hassoun LA, Sivamani RK. A systematic review of lactoferrin use in dermatology. Crit Rev Food Sci Nutr. 2017 Nov 22;57(17):3632–3639. doi: 10.1080/10408398.2015.1137859.

8. Nikolaev A.A., Sukharev A.E. Lactoferrin and its role in reproduction (literature review). Russian Journal of Human Reproduction. 2005. Vol. 21, No.6, pp. 33–38.

9. Ploskonos M.V., Syatkin S.P., Neborak E.V., Hilal A., Sungrapova K.Yu., Sokuyev R., Blagonravov М., Korshunova A., Teren-tyev A.A. Polyamine analogues of propanediamine series inhibit prostate tumor cell growth and activate polyamine catabolic pathway. Anticancer Research, vol. 40 no. 3, 2020, Р. 1437–1441. doi:10.21873/anti-canres.14085

10. Kell DB, Heyden EL, Pretorius E. The biol-ogy of lactoferrin, an iron-binding protein that can help defend against viruses and bacteria. Front Immunol. 2020 May 28;11:1221. doi: 10.3389/fim-mu.2020.01221.

11. Morgenthau A, Livingstone M, Adamiak P, Schryvers AB. The role of lactoferrin binding pro-tein B in mediating protection against human lactofer-ricin. Biochem Cell Biol. 2012 Jun;90(3):417–23. doi: 10.1139/o11-074.

12. Morgenthau A, Pogoutse A, Adamiak P, Moraes TF, Schryvers AB. Bacterial receptors for host transferrin and lactoferrin: molecular mecha-nisms and role in host-microbe interactions. Future Microbiol. 2013 Dec;8(12):1575–85. doi: 10.2217/fmb.13.125.

13. Neumann W, Hadley RC, Nolan EM. Transition metals at the host-pathogen interface: how Neisseria exploit human metalloproteins for acquiring iron and zinc. Essays Biochem. 2017 May 9;61(2):211–223. doi: 10.1042/EBC20160084.

14. Pammi M, Suresh G. Enteral lactoferrin supplemen-tation for prevention of sepsis and necrotizing entero-colitis in preterm infants. Cochrane Database Syst Rev. 2017 Jun 28;6(6):CD007137. doi: 10.1002/14651858.CD007137

15. Shi Y, Kong W, Nakayama K. Human lactoferrin binds and removes the hemoglobin receptor protein of the periodontopathogen Porphyromonas gingiva-lis. J Biol Chem. 2000 Sep 29;275(39):30002–8. doi: 10.1074/jbc.M001518200.

16. Telang S. Lactoferrin: A Critical Player in Neonatal Host Defense. Nutrients. 2018 Sep 4;10(9):1228. doi: 10.3390/nu10091228.

17. Ulvatne H, Samuelsen Ø, Haukland HH, Krämer M, Vorland LH. Lactoferricin B inhibits bacterial macromolecular synthesis in Escherichia coli and Bacillus subtilis. FEMS Microbiol Lett. 2004 Aug 15;237(2):377–84. doi: 10.1016/j.femsle.2004.07.001.

18 Pham J.V, Yilma, M.A., Feliz, A. et al. Review of the Microbial Production of Bioactive Natural Prod-ucts and Biologics Front. Microbiol., 20 June 2019 https://doi.org/10.3389/fmicb.2019.01404



PERSONALIZED FSI-MODELING OF THE AORTIC BULB AND ARCH TO PREDICT ITS MECHANICAL BEHAVIOR AND ASSESS THE LOADS DURING THE CARDIAC CYCLE

A B S T R A C T — The interaction of the blood flow with the aorta is a complex dynamic event described in biomechanics as the Fluid-structure interaction. In this study we’ve developed a method for creation of a personalized 3D dynamic model of the aortic bulb and arch for the prediction of its mechanical behavior using FSI-analysis. We found that the accuracy of predicting geometric aortic deformities based on FSI modeling is on average 92%.

K E Y W O R D S — aorta, aortic valve, hemodynamics, FSI-modeling.

Artur Ovsepyan2,4 , Alexander Smirnov1,2 ,Sergey Dydykin3 , Yuriy Vasil’ev3 ,Evgeniy Trunin6 , Olga Shatunova5,Aleksandr Aleksandrov6 , Angelina Ostyakova6 ,Andrey Utkin7

1 Moscow State Regional University, Moscow;2 St. Petersburg State University, Laboratory of Biomechanical Analysis. St. Petersburg;3 I.M. Sechenov First Moscow State Medical University (Sechenov University), Moscow;4 St. Petersburg State Electrotechnical University "LETI", St. Petersburg;5 N.A. Semashko City Multidisciplinary Hospital No. 38, Pushkin;6 I.I. Mechnikov North-Western State University, St. Petersburg;7 Kazan State Medical University, Kazan, Russia

[email protected]

R E L E V A N C EThe interaction of the blood flow with the aorta

is a complex dynamic event described in biomechan-ics as the interaction of a fluid and a deformable body (Fluid-structure interaction or FSI). The existing methods of in vivo visualization and quantitative analysis in silico allow us to model these processes in order to study the pathogenesis of diseases of the cardiovascular system, predict risks and plan surgical interventions [1]. The FSI method is widely used for numerical modeling of the blood-vessel interaction [2, 3, 4]. It combines the methods of computational fluid dynamics and structural (dynamic) analysis. Today, FSI is widely used in predicting the risks of

Received 19 March 2021 Received in revised form 20 April 2021; Accepted 8 Mai 2021

aneurysms and their ruptures. Previously, no attempts have been made to study changes in the biomechani-cal properties of the aorta in aortic valve stenosis using personalized FSI modeling based on MSCT data.

Aim:to create a personalized FSI model of the patient's aortic segment and then to evaluate its mechanical behavior during the cardiac cycle.

M A T E R I A L S A N D M E T H O D SFunctional MSCT-coronarography data were

used to model the aortic segment. Based on these data, in the InobitekDicom Viewer software, a multiplanar reconstruction of the zone of interest was generated, including the anatomical structures of the heart and aorta. For the aortic valve, the ascending aorta, and the aortic arch, routine segmentation was performed in three projections. From the resulting set of contours, a three-dimensional model was created, which was then converted into a polygonal STL model (Fig. 1).

Fig. 1. STL model of the zone of interest

Editing and reverse engineering of the STL model was performed in the SolidWorks software. The aortic wall and its root were generated using the SW ScanTo3D utility. The valves and commissures of the aortic valve were modeled using standard SW tools.


http://dx.doi.org/10.35630/2199-885X/2021/11/2/3

https://orcid.org/0000-0002-4050-214X

https://orcid.org/0000-0002-2661-3759

https://orcid.org/0000-0002-1273-0356

https://orcid.org/0000-0003-3541-6068

https://orcid.org/0000-0002-2452-0321

https://orcid.org/0000-0003-0012-8429

https://orcid.org/0000-0002-1437-8974

https://orcid.org/0000-0002-6343-4581

http://dx.doi.org/10.35630/2199-885X/2021/11/2/3


The CAD model of the zone includes the structures of the aorta, the fibrous ring, the valves, commissures and sinuses of the Valsalva (Fig. 2).

junction of the aorta) and at the exit of the blood flow (in the area of the brachiocephalic trunk, the left com-mon carotid artery and the left subclavian artery) in 4 nodes located in the centers of the above areas.

Each node is connected by the condition of the continuum distribution with the elements of the aortic wall in the planes of sections at the entrance and exit, without limiting the degrees of freedom of the con-nected elements. In the area of the border of the arch and the descending part of the aorta, the model is fixed in the area of the arterial ligament with the condition of the distribution of the continuum with the elements of the aortic wall in the cross-sectional plane without limiting the degrees of freedom of the connected ele-ments.

The blood flow is specified as laminar, the flow rate at the peak of the systole is v = 1.2 m/s (Fig. 4) [5], the flow input is the entire area in the plane of the sino-tubular junction of the aorta, the output is the areas of the brachiocephalic trunk, the left common carotid artery, the left subclavian artery, the border of the arch and the descending aorta (outlet pressure P = 0).

The connection condition (FSI) between the aorta and the fluid was set by the Fluid-Structure Co-simulation boundary parameter. This parameter describes the interaction between the Computational Fluid Dynamics (CFD) module and the Abaqus CAE Implicit Dynamic Analysis (Dynamic/Implicit) module. The choice of the Dynamic/Implicit module is based on the hypothesis of small deformations in the simulated segment, which eliminates the need to use adaptive meshes.

For the subsequent formulation of the FSI prob-lem, the flow region was modeled. The final grid of finite elements for aortic structures consists of 32109 tetrahedral elements, for the flow region (blood) — 34999 tetrahedral elements. To model the anisotropic properties of aortic structures, we used the Holzapfel-Gasser-Ogden (HGO) anisotropic material model. To evaluate the proposed model, an FSI analysis of the aortic segment, including the ascending section and the aortic arch, was performed during systole (nor-mal). The results of the analysis were compared with the MSCT data and analyzed by the expert control method. In this paper, the model adequacy criteria were taken as the deformed states of the FSI model, which were compared with the displacements on the MSCT at the corresponding moments of the systole. Aortic valve stenosis was modeled by changing the area and peak flow rate during systole at the model inlet.

R E S U L T SFor the validation model, Mises displacement,

pressure, and stress plots were obtained for the aortic

Fig. 2. Aortic valve model: 1 — bottom view, 2 — top view, 3 — split view, 4 — full CAD model

1 2

3

4

Preprocessing of the CAD model for FSI analysis was performed in HyperMesh software. The calculated KE-grid was generated taking into account the ana-tomical and morphological features of the aortic wall.

The resulting CE model was imported as an orphan grid in Abaqus CAE to set the mechanical properties of the materials and set the boundary con-ditions for the biomechanical analysis of the model. To optimize the computational load, we introduced the following boundary conditions: the model is fixed (Fig. 3) at the entrance (the plane of the sinotubular

Fig. 3. Fixed geometry: pink — connected elements of the aortic wall without restrictions of degrees of freedom, the intersection of yellow lines — a node with a fixed geometry



Fig. 4. Graph of changes in the flow rate during the cardiac cycle. The simulated area is 0.23 seconds from the beginning to the peak of the systole

wall. Changes in the blood flow characteristics in the model are described. All initial conditions were mod-eled for systole at time t = 0 in accordance with the MSCT data at the same time. The FSI analysis made it possible to predict the behavior of the aortic wall dur-ing a systole lasting 0.23 seconds. The prediction result (displacement) was compared with the MSCT at time t = 0.1 s, and t = 0.23 c corresponding to the peak of systole (Fig. 5).

Fig. 5. Plots of movements in meters (line 2) at the cor-responding moment

We investigated the geometric deviations of the FSI simulation from the MSCT data by superimpos-ing and analyzing the STL meshes extracted from the solver output file at time t = 0, t = 0.1 c, t = 0.23 c, on

the 3D reconstruction of the MSCT for the corre-sponding time points. The overlay of each STL-grid was carried out by means of rigid registration of images based on the intensity (Fig. 6).

The IoU coefficient for the volume model was calculated as the arithmetic mean IoU of all 2D im-ages. The maximum value of the coefficient for the two forecasts of the FSI model is 0.96, and the minimum value is 0.73. The average IoU value for the two models

was 0.92. Figure 7 shows the Mises stress plots (1), the pressure distribution along the aortic wall (2), and the flow characteristics at the peak of systole t = 0.23 s (3) in normal, predicted by FSI modeling.



C O N C L U S I O NWe established that the accuracy of predicting

geometric aortic deformities based on FSI modeling is on average 92%. The results of modeling using the proposed methods do not contradict the previously obtained data in silico. The predicted deformed states correspond well to the in vivo data.

R E F E R E N C E S1. Heydarov N. A., Gainullova K. S., Drygina

O. S. Computer methods for modeling blood flow regarding the problems of cardiology and cardiac sur-gery //Complex problems of cardiovascular diseases. - 2018. - Vol. 7. - No. 2.

2. Bahraseman H. G. et al. Fluid-Structure Interac-tion modeling of aortic valve stenosis at different heart rates //Acta of bioengineering and biomechanics. – 2016. – Т. 18. – No. 3. DOI: 10.5277/ABB-00429-2015-03

3. Mao W. et al. Fully-coupled fluid-structure interac-tion simulation of the aortic and mitral valves in a realistic 3D left ventricle model //PloS one. – 2017. –

Fig. 6. MSCT-FSI combined 3D reconstructions. The deformed STL meshes are highlighted in pink. The anatomical structures of MSCT reconstruction are encoded by a soft tissue imaging filter

Fig. 7. Prediction of hemodynamics and mechanical behavior of the aorta in normal conditions

Т. 12. – No. 9. – С. e0184729. https://doi.org/10.1371/journal.pone.0184729

4. Gilmanov A. et al. Image-guided fluid-structure interaction simulation of transvalvular hemodynamics: Quantifying the effects of varying aortic valve leaflet thickness //Fluids. – 2019. – Т. 4. – No. 3. – С. 119. https://doi.org/10.3390/fluids4030119

5. Spühler J. H. et al. 3D fluid-structure interaction simulation of aortic valves using a unified con-tinuum ALE FEM model //Frontiers in physiology. – 2018. – Т. 9. – С. 363. https://doi.org/10.3389/fphys.2018.00363



EFFECT OF LEAD ON STRUCTURAL CHANGES IN LIVER OF WISTAR RATS UNDER THE CONDITIONS OF ACUTE EXPERIMENT

A B S T R A C T — The purpose of this study was to evaluate the structural transformations of Wistar rats liver after oral administration of lead salts. Under the conditions of acute experiment during 5 days the Wistar rats were orally administered the lead acetate solution in the amount of 3 mg/kg. Histological studies were carried out at OSMU upon the completion of the experiment. Effect of toxic doses of lead causes symptoms of both steatosis and hydropic degeneration of the liver. It is assumed that the appearance of hepatic steatosis reflects the reaction of hepatocytes to hemic hypoxia caused by the action of lead, while the signs of hydropic degeneration expressed in varying degrees reflect its direct toxic effect.

K E Y W O R D S — lead, liver, hepatocytes, hydropic degeneration, hepatic steatosis.

Oleg Zayko1 , Vadim Astashov1 ,Anna Sindireva2 , Karina Basnakyan1 ,Arina Lukyanchikova

1 RUDN University, Moscow, Russia 2 Tyumen State University, Tyumen, Russia

[email protected]

R E L E V A N C EIn recent decades, heavy metals are increasingly

frequently named as the most dangerous environmen-tal pollutants. Their migration and redistribution in the components of ecosystems depend both on the whole complex of natural factors, and the intensity and nature of technogenesis. Lead is not a vital element. It is toxic and belongs to danger class I. Its inorganic compounds interfere with the metabolism and are the enzymes inhibitors (like most of the heavy metals). One of the most insidious consequences of the action of inorganic lead compounds is its ability to accumu-late in a number of organs, including the liver, and to be a constant source of poisoning for a long time. The main source from which lead enters the body, is the food, and also the inhaled air plays the important role. Absorption in the gastrointestinal tract is up to a 5–10% in adults, and 50% in children. Thus, there is a need to assess the toxic effect of lead.

Received 22 March 2021; Received in revised form 1 May 2021; Accepted 5 May 2021

The purpose of this study: to evaluate the structural transformations in the liver of Wistar rats after oral administration of lead salts.

M E T H O D S20 male Wistar rats aged 4–5 months, weigh-

ing 200–220 g were divided into an experimental and control group — 10 rats in each group. The rats were maintained in standard cages under regular daily al-ternation of light and darkness, temperature 20–22° C and free access to water and food. The studies were carried out in in compliance with the CIOMS-ICLAS International Guiding Principles for Biomedical Re-search Involving Animals.

Under the conditions of acute experiment during 5 days the Wistar rats were orally administered the lead acetate solution in the amount of 3 mg/kg. Histologi-cal studies were carried out at OSMU upon the com-pletion of the experiment. The cut and oriented liver acinuses were fixed in 10% neutral buffered formalin (pH = 7.2–7.4). They were dehydrated and embed-ded in paraffin according to conventional technique. 4–5 microns thick paraffin sections were stained with hematoxylin and eosin. Ziehl-Neelsen method (detection of fuchsinophile acid resistant intranuclear inclusions) by Lily’s formulation (1969) was used to identify lead pellets in a complex with protein deposits in the form of intranuclear inclusions in the cells of renal tubule. Sections were stained with carbol fuchsin (dissolve 25 g of phenol in 50 ml alcohol, then in 5 g of basic fuchsin, fill with distilled water to 500 ml) over a fire until the vapor (4–5 minutes), then the glass was cooled down and colorant drained. Rinsing was performed in running water with differentiation in a 5% solution of sulfuric acid, followed by counterstain-ing with methylene blue. Pearls reaction was used to identify hemosiderin.

R E S U L T SThe following data were obtained after the

analysis of liver micropreparations of the experimental animals. During an acute experiment in lead poison-ing conditions, the signs of hydropic degeneration in the hepatic tissue were observed from the very first day of the experiment. Predominant changes were ob-served in centrolobular localization hepatocytes, while


http://dx.doi.org/10.35630/2199-885X/2021/11/2/4

https://orcid.org/0000-0002-9324-3591

https://orcid.org/0000-0003-2846-1944

https://orcid.org/0000-0001-8596-7584

https://orcid.org/0000-0001-6878-0312

https://orcid.org/0000-0002-2810-3146

http://dx.doi.org/10.35630/2199-885X/2021/11/2/4


proteinosis, recognizable in the light microscope by the irregular cytoplasm (crumpled up paper symptom) was represented unevenly, in the form of peculiar nests. Plethora of central veins and expansion of adjacent sinusoids were observed, and the said changes were already significantly expressed at day 3 or more of the experiment (fig. 1).

Fig. 3. Hepatic tissue amid acute experiment of lead poisoning. Clustering of Kupffer cells in congested dilated sinusoids, regenerative transforma-tion of hepatocytes. Haematoxylin and eosin staining. ×900

The lumen of the sinusoids and perisinusoidal space was marked with the appearance of hepatic mac-rophages — Kupffer cells as a rule arranged in groups up to 5–8 cells (fig. 3).

Signs of proteinosis were characterized by a greater area of distribution covering even the peripher-al parts of the acinus. The dystrophy in some hepato-cytes of centrolobular localization was expressed up to a balloon type, with the cells with signs of colliquative necrosis (fig. 2).

Fig. 1. Lead poisoning. Dilatation and congestion of central segments of sinusoids. Haematoxylin and eosin staining. ×270

Fig. 2. Acute lead poisoning. Dilatation and congestion of sinusoids. Coagulative necrosis of hepatocytes, mitoses. Haematoxylin and eosin staining. ×900

By the end of the experiment the majority of the formulations had the signs of not only proteinosis, but also steatosis with small-drop character and represent-ed diffusely in liver acinus (fig. 4).

Fig.4. Lead poisoning. Disseminated small-droplet fatty degeneration of hepatocytes. Coagulative necrosis of certain cells. Haematoxylin and eosin staining. ×900

Hepatocytes with regenerative changes, hyper-chronia of cores with increased size, appearance of binuclear hepatocytes were also observed, mitosis was rarely observed.



C O N C L U S I O NThe effect of toxic doses of lead results in symp-

toms of both hepatic steatosis and hydropic degenera-tion of the liver. It is assumed that the appearance of steatosis reflects the reaction of hepatocytes to hemic hypoxia caused by the action of lead, while the signs of hydropic degeneration is expressed in varying degrees reflect its direct toxic effect.

R E F E R E N C E S1. Stuart J Forbes, Philip N Newsome «Liver

regeneration – mechanisms and models to clinical application» // Natural Reviews. Gastroenterology & Hepatology. – 2016 Aug;13. – №8. – p.473–485 DOI: 10.1038 / nrgastro.2016.97.

2. Jorge Vizuete, Marcos Pérez-López, María Prado Míguez-Santiyán, David Hernández-Moreno «Mercury (Hg), Lead (Pb), Cadmium (Cd),

Selenium (Se), and Arsenic (As) in Liver, Kidney, and Feathers of Gulls: A Review» // Reviews Environmen-tal contamination and toxicology. – 2019. P. 85–146 DOI: 10.1007 / 398_2018_16.

3. Alan N Hunt, Anagha Malur, Tual Mon-fort, Pavlos Lagoudakis, Sumeet Mahajan, Anthony D Postle, Mary Jane Thomassen «Hepatic Steatosis Accompanies Pulmonary Alveolar Proteinosis» // American journal of Respiratory Cell and Molecular Biology. – 2017. – №4. – p.448–458 DOI:10.1165/rcmb.2016-0242OC.

4. Ju-Wook Lee, Hoon Choi, Un-Ki Hwang, Ju-Chan Kang, Yue Jai Kang, Kwang Il Kim, Jun-Hwan Kim «Toxic effects of lead exposure on bioaccumulation, oxidative stress, neurotoxicity, and immune responses in fish: A review» // Environ-mental Toxicology and Pharmacology. – 2019. – p. 101–108 DOI: 10.1016/j.etap.2019.03.010



INFLUENCE OF COPPER COMPLEXES [CU(PTA)4[BF4] AND CU(II)2(3,5-DIPS)4(H2O)3 ON THE ORGANISM OF RATS IRRADIATED WITH RADIOISOTOPE TECHNETIUM

A B S T R A C T — One of the priority tasks of modern radiobiology is the search for new, effective radioprotective compounds. In this area, metal-organic complexes with high antioxidant activity are of particular interest.In order to study the possible beneficial radioprotective effect of copper complex compounds [Cu(PTA)4[BF4] and Cu(II)2(3,5-DIPS)4(H2O)3 on an irradiated organism, we studied cytogenetic parameters in 4 groups of experimental animals: intact animals, animals exposed to the radioisotope technetium (Tc) — "pure irradiation", animals with "irradiation + compound [Cu(PTA)4[BF4]", and animals with "irradiation + compound [Cu(II)2( 3,5-DIPS)4(H2O)3".The survival rate and cytogenetic parameters were studied: mitotic index (MI), chromosomal aberrations (CA) and polyploid cells (PC) in the bone marrow cells of the femur.The survival rate in the first and third groups was 100%, in the second group — 40%, and in the fourth — 80%. The dynamics of survival was described by regression curves and equations, which make it possible, using extrapolation, to determine the change in the percentage of survival in the long term of the experiment and to predict the further outcome of the experiment.When analyzing the results in 4 groups of animals, we found a significant difference in cytogenetic parameters between these groups. Thus, for all 3 indicators, a significant difference is observed between intact and irradiated animals, i.e. these parameters can be considered as markers of Tc exposure. In terms of the mitotic index (proliferative activity), a significant difference was found in the irradiated compared with the groups: "irradiation + [Cu(PTA)4[BF4]" and "irradiation + Cu(II)2(3,5-DIPS)4(H2O)3", which indicates the radioprotective property of both compounds. By the number of polyploid cells, a significant difference was found between the groups: "pure irradiation" and "irradiation + Cu(II)2(3,5-DIPS)4(H2O)3", which also proves the beneficial effect of this compound.Multiregression analysis of cytogenetic parameters along with standard statistical methods confirmed the highest efficiency of [Cu(PTA)4[BF4] relative to Cu(II)2(3,5-DIPS)4(H2O)3.

K E Y W O R D S — radiation, chromosomal aberrations, polyploid cells, mitotic index, survival.

Anahit Karapetyan1 , Ashot Dallakyan1,Marina Porchia2 , Carlo Santini3 ,Gayane Khachatryan1 , Vahan Grigoryan1

1 National Burn Center, Yerevan, Armenia 2 ICMATE, National Research Council (CNR), Padua, Italy 3 School of Science and Technology, University of Camerino, Italy

[email protected]

I N T R O D U C T I O NIt is known that the main initiating event after ir-

radiation of an organism is DNA damage, on the basis of which chromosome destabilization is considered one of the first and direct signs of the effect of ioniz-ing radiation (IR) on a cell [1–4]. Radiation-induced damage to the karyotype is an important indicator both for biological indication of the severity of radia-tion injuries and for predicting the development of long-term adverse effects of IR.

One of the priority tasks of modern radiobiology is the search for new, effective radioprotective com-pounds. In this area, metal-organic complexes with high antioxidant activity are of particular interest.

The ability to protect the body from the damag-ing effects of ionizing radiation in such complexes were noted both in the scientific works of other authors [5, 6] and in our early works [7–10].

Aim: to study the possible beneficial radioprotective effect of copper complex compounds [Cu(PTA)4[BF4] and Cu(II)2(3,5-DIPS)4(H2O)3 on an irradiated organism using cytogenetic parameters.

M A T E R I A L S A N D M E T H O D SIn order to study the possible beneficial ra-

dioprotective effect of copper complex compounds [Cu(PTA)4[BF4] and Cu(II)2(3,5-DIPS)4(H2O)3 on an irradiated organism, we studied cytogenetic param-eters in 4 groups of experimental animals (white, out-bred, sexually mature male rats with an average weight of 180 g,; 10 rats in each group).

Received 8 May 2921; Received in revised form 30 May 2021; Accepted 2 June 2021

The animals used in our research were maintained in compliance with European Union Legislation (Directive 2010/63/EU, amended by Regulation (EU) 2019/1010).

The first group included intact animals. The sec-ond group consisted of animals exposed to the radioi-


http://dx.doi.org/10.35630/2199-885X/2021/11/2/5

https://orcid.org/0000-0002-3318-3881

https://orcid.org/0000-0003-0435-6576

https://orcid.org/0000-0002-3942-1713

https://orcid.org/0000-0001-5250-8365

http://dx.doi.org/10.35630/2199-885X/2021/11/2/5


sotope technetium (Tc), which were injected intraperi-toneally with an isotope with an activity of 4.8 mCi in a volume of 2 ml — "pure irradiation". The third group consisted of animals that were injected intraperitoneal-ly with a copper complex [Cu(PTA)4[BF4] at a dose of 50 mg/kg in a volume of 2 ml one hour before the administration of the Tc isotope («irradiation + cop-per compound [Cu (PTA)4[BF4])»). The fourth group involved animals that, before irradiation, received the compound Cu(II)2(3,5-DIPS)4(H2O)3 at a dose of 50 mg/ kg in a volume of 1 ml.

We studied the survival rate and cytogenetic parameters at the metaphase stage of the mitotic cycle (according to the Ford-Wallam method [11], the following were determined: the mitotic index (MI), chromosomal aberrations (CA) and polyploid cells (PC) in the bone marrow cells of the femur (counting in 1000 cells in each). The cytogenetic examination included the analysis of chromosomes using a J.B. Carnoy fixative and with Giemsa stain. The obtained cytogenetic preparations were analyzed according to the generally accepted method of cytogenetic analysis of bone marrow cells (BMC) of white rats according to G. McGregor [12] under a microscope at 900–1400 times magnification.

Data analysis was carried out using a number of specialized statistical packages: Statsoft and SPSS-10.0. We used regression, multi-regression and correlation methods of analysis [13, 14].

R E S U L T SThe experiment lasted 30 days. The survival rate

of the second group was 40%. In the third group, with the injection of the compound [Cu(PTA)4[BF4], the survival rate was 100%, and in the fourth — 80%. The dynamics of survival was described by regres-sion curves and equations (where y1 is survival under pure irradiation, y2 is when «irradiated + injection of Cu(II)2(3,5-DIPS)4(H2O)3» and y3 is when «irradi-ated + injection [Cu(PTA)4[BF4])» shown in Fig. 1, which make it possible, using extrapolation, to deter-mine the change in the percentage of survival in the long term of the experiment and to predict the further outcome of the experiment.

Analyzing the karyotype and proliferative activity of the above cells, we obtained the cytogenetic indica-tors of these groups, the results of which are shown in the Table 1. Only reliable values of changes in cytoge-netic parameters are given.

The types of karyotype disorders (polyploid cell and double fragment) detected after irradiation of animals with technetium are shown in Fig. 2.

When analyzing the results of the animal groups "pure irradiation", "irradiation + [Cu(PTA)4[BF4]" and

Fig. 1. Survival of the groups "pure irradiation", "irradiation + [Cu(PTA)4[BF4]" and "irradiation + Cu(II)2(3,5-DIPS)4(H2O)3"

"irradiation + Cu(II)2(3,5-DIPS)4(H2O)3", we found a significant difference in cytogenetic parameters between these groups. Thus, for all 3 indicators, a significant difference is observed between intact and irradiated animals, i.e. these parameters can be consid-ered as markers of Tc exposure. In terms of the mitotic index (proliferative activity), a significant difference was found in the irradiated compared with the groups: "irradiation + [Cu(PTA)4[BF4]" and "irradiation + Cu(II)2(3,5-DIPS)4(H2O)3", which indicates the radio-protective property of both compounds. According to the number of polyploid cells, a significant difference was found between the groups: "pure irradiation" and "irradiation + Cu(II)2(3,5-DIPS)4(H2O)3", which also proves the beneficial effect of this compound.

Fig. 3 shows the results of multi-regression dependences of the mutual influence of cytogenetic parameters upon injection of [Cu(PTA)4[BF4] (a) and Cu(II)2(3,5-DIPS)4(H2O)3 (b). In the formulas shown in the upper part of the figure (a) and (b) x is the coefficient characterizing the change in cy-togenetic parameters under normal conditions, y is under the influence of Technetium, z1 is under the influence of Tc + [Cu(PTA)4[BF4] (a) , and z2 — un-der the influence of Cu(II)2(3,5-DIPS)4(H2O)3 (b). Multiregression analysis of cytogenetic parameters along with standard statistical methods confirmed the highest efficiency of [Cu(PTA)4[BF4] relative to Cu(II)2(3,5-DIPS)4(H2O)3.

The research results indicate the need to continue work in the direction of searching for drugs that have a therapeutic effect in radiation injuries.

R E F E R E N C E S1. Mazurik V.K., Mikhailov V.F. Radiation-induced

genome instability: phenomenon, molecular mecha-nisms, pathogenetic significance. Radiobiology and radioecology, 2001, v.41, 3, p.272–289



2. Alberts B., Johnson A., Lewis J., Raff M., Rob-erts K., Walter P. Molecular Biology of the Cell. 5th edition., 2007.

3. Khvostunov I.K., Pyatenko V.S., Shepel N.N., Korovchuk O.N., Golub E.V., Zhironkina A.S., Khvostunova T.I., Lychagin A.A. Analysis of chromosomal aberrations in mammalian cells under the influence of various types of ionizing radiation, Radia-tion and Risk, 2013, v. 22, N 4, Obninsk, RF, p.43–59

4. Minina V.I. Genetic polymorphism and chromosom-al aberrations induced by radiation, Siberian Medical Journal, 2012, v. 110, N 3, Irkutsk, RF, p.5–7, ISSN: 1815-7572

5. Malakyan M.H., Bajinyan S.A., Matosyan V.A., Tonoyan V.J, Babayan K.N., Yegiazaryan D.E. Amino acid Schiff bases and their Cu (II) chelates are new highly effective radioprotectors. Bul-letin of the Russian Military Medical Academy, 2008, N 3 (23), p.219

groupsindicators

Norm(group 1)

Tc(group 2)

Тс+ [Cu(PTA)4[BF4] (group 3)

Тс+ Cu(II)2(3,5-DIPS)4(H2O)3(group 4)

Mitotic index (MI) ‰ 20,1±2,8 10,9±0,350,01<pn2<0,02

14,2±0,960,01<p23<0,02

17,2±1,90,002<p24<0,01

Chromosomal aberra-tions (ChA) % 3,0±0,22 6,2±0,5

pn2<0,0014,8±0,420,002<pn3<0,01

4,8±0,50,002<pn4<0,01

Polyploid cells (PC) % 0.001±0.0001 3,5±0,44pn2<0,05

3,4±0,47pn3<0,05

2,0±0,2pn4<0,05 0.01<p24<0.02

Table 1. Cytogenetic indices in 4 groups: "normal", "pure irradiation", "irradiation + [Cu(PTA)4[BF4]" and "irradia-tion + Cu(II)2(3,5-DIPS)4(H2O)3" on the 30th day of the experiment

pn2

— when comparing the indicators of the second group and the group of intact animalsp

n3 — when comparing the indicators

of the third group and the group of intact animalsp

n4 — when comparing the indicators

of the fourth group and the group of intact animalsp

23 — when comparing the indicators

of the second and third groupsp

24 — when comparing the indicators

of the second and fourth groups

Fig. 2. Polyploid cell (a) and double fragment (b) 45 days after exposure to technetium isotope

Fig. 3. Interdependence of cytogenetic parameters upon injection of [Cu(PTA)4[BF4] (a) and Cu (II)2(3,5-DIPS)4(H2O)3 (b)

a b

6. Malakyan M.G., Bajinyan S.A., Matosyan V.A., Babayan K.N., Tonoyan V.J, Yeghiazaryan D.E., Vardevanyan L.A. Investigation of the radioprotective and antiradical activity of Schiff bases, nicotinaldehyde and L-amino acid derivatives. In the Collection of Materials of the International Scien-tific and Practical Interdisciplinary Seminar "New Technologies in Medicine and Experimental Biology", 2007, Bangkok-Pattaya, p. 44–46.

7. Dallakyan A.M., Petrosyan J.G., Harutyun-yan N.K., Karapetyan A.G. Changes in peripheral blood and cytogenetic parameters when using com-pounds of copper-organic complexes: [Cu(PTA)4BF4 and [Cu(PCN)(HBP23)]BF4 before irradiation, Medical Science of Armenia, Yerevan, 2019, v. LIX, N 1, p.76–86

8. Karapetyan A.G., Santini K., Malakyan M.G. Evaluation of the duration and effectiveness of the radioprotective action of the new synthesized com-pounds. Medical Science of Armenia, Yerevan, 2019, v. LIX, N 2, p.12–19

9. Karapetyan A.G., Dallakyan A.M., Tonoyan V.D. Cytogenetic assessment of the radioprotective properties of Cu and Mn chelates of Schiff bases of Nicotinyl-L-amino acids. Medical Science of Armenia, Yerevan, 2019, v. LIX, N 3, p. 34–40

10. Oganesyan N.M., Santini C., Karapetyan A.G., Tisato F., Porchia M., Dallakyan A.M. et al. Changes in peripheral blood and cytogenetic indicators during the application of compounds of the copper-organic complexes to irradiation, Archiv Euromedica, 2019, v. 9, N 2, p. 68–71 https://doi.org/10.35630/2199-885X/2019/9/2/68

11. Ford E., Wollam D. A Study of the Mitotic Chro-mosomes of Mice of the Strong a Line, Experimental Cell Research v. 32, p. 320–326 DOI: 10.1016/0014-4827(63)90106-x

12. McGregor G., Varley J. Methods of working with animal chromosomes. M., "Mir" 1986.

13. Vukolov E.A. Fundamentals of Statistical Analysis. Workshop on statistical methods and operations research using Statistica and Excel packages, Moscow, Forum-Infra, M, 2004, – 464 p.

14. Gmurman V.E. Probability theory and mathematical statistics: Textbook for universities, M: Higher school, 2004, – 479 p.



EVALUATION OF THE CORTICOSTEROID RECEPTORS’ LEVEL IN THE KIDNEYS OF RATS AFTER SYSTEMIC ISCHEMIA-REPERFUSION

A B S T R A C T — Background: With disturbance of the systemic blood supply, the body experiences hypoxia and stress. Under stress of any etiology, there is a non-specific rearrangement of physiological and biochemical processes. These processes occur under the influence of corticosteroid hormones. Aim: To determine the level of corticosteroid receptors in the kidneys of rats at different times after systemic ischemia-reperfusion. Methods: The study included 80 male white rats. All the animals were divided into 2 groups. A model of systemic ischemia-reperfusion was created in the main group (n=70). Further, on 1, 3, 5, 7, 14, 21 and for 35 days, we determined the level of gluco - and mineralocorticoid receptors in the kidneys. Results: In the animals of the main group, we observed a short-term period (during the first 3 days) of a decrease in the content of both glucorticoid (p<0.05) and mineralocorticoid (p<0.01) receptors. The dynamics of recovery of the level of corticosteroid receptors was 3 times faster than that of mineralocorticoid receptors. Conclusions: The dynamics of corticosteroid receptors’ level in the kidneys of rats after ischemia caused by an arrest of systemic circulation show the recovery time after ischemia-reperfusion injury, which ensures the stability of an individual to hypoxia.

K E Y W O R D S — glucocorticoid receptors, mineralocorticoid receptors, resistance to hypoxia, ischemia-reperfusion injury.

Gulnar Bayburina1 , Elena Nurgaleeva1,Ajgul Samigullina1 , Ekaterina Farshatova1, Eduard Agletdinov1, Tatyana Tarasova2 ,Larisa Mosina2, Sergey Ryzhakin3, Dalila Khaydar3

1 Bashkir State Medical University, Ufa, Bashkortostan. Russia2 National Research Ogarev Mordovia State University, Saransk, Russia, 3 Peoples Friendship University of Russia (RUDN University), Moscow, Russia

[email protected]

occurs, which is necessary to ensure the coordinated functioning of all body systems and adaptation to the pathogenic factor [1].

In case of disruption of the systemic blood supply, the effects of corticosteroid hormones can be signifi-cantly distorted due to changes in the sensitivity of target tissue receptors and disorders in the processes of hormone biotransformation, which can cause serious maladaptive consequences [2].

The kidney is an organ that is very sensitive to hemodynamic disorders and ischemia [3]. Therefore, the study of the reaction of resistance to hypoxia after ischemia-reperfusion disorders is an urgent issue.

Aim: To determine the level of corticosteroid receptors

in the kidneys of rats at different times after systemic ischemia-reperfusion.

M E T H O D SThe study was experimental in type and included

80 mature male white rats. The study was carried out in accordance with the ethical standards for the treat-ment of animals adopted by the European Convention for the Protection of Vertebrate Animals for Research and Other Scientific Purposes, the Federation of European Associations for the Laboratory Animal Sci-ence and the International Council for the Laboratory Animal Science.

All animals were divided into two groups:— main group (n=70; 10 rats for each stage of

observation). In this group, we created a model of systemic ischemia-reperfusion.

— control group (n=10).

At first, we gave ether anesthesia to all the animals of both groups. Then, we simulated a 5-minute stop of systemic circulation by clamping the vascular bundle of the heart intrathoracically under ether anesthesia to the rats of the main group. Further, we carried out resuscitation measures in these animals: external heart massage and artificial lung ventilation. In the control group of rats after ether anesthesia, the circulatory ar-rest was not reproduced.

We evaluated the response of resistance to hypoxia after systemic ischemia-reperfusion based on

Received 14 May 2921; Received in revised form 3 June 2021; Accepted 7 June 2021

I N T R O D U C T I O N The complex of protective and compensatory

reactions that occur under the influence of extreme hypoxia primarily involves the activation of the hypothalamic-pituitary-adrenal system. It is known that under the influence of corticosteroid hormones (CSH) under stress of any etiology, a nonspecific rear-rangement of physiological and metabolic processes


http://dx.doi.org/10.35630/2199-885X/2021/11/2/6

https://orcid.org/0000-0002-4741-6295

https://orcid.org/0000-0001-9026-7259

http://orcid.org/0000-0001-9745-9739

https://orcid.org/0000-0002-5490-1037

http://dx.doi.org/10.35630/2199-885X/2021/11/2/6


the level of gluco- and mineralocorticoid receptors in the kidney tissue homogenates. The level of receptors was determined on the analyzer "StatFox 2100" by the method of enzyme immunoassay, using standard test kits ELISA Kit (China) from Cloud-Clone Corp (USA). We studied the level of corticosteroid recep-tors on 1, 3, 5, 7, 14, 21 and 35 days.

Statistical processing of the material was car-ried out using the program "STATISTICA 7.0". The confidence of differences between quantitative indica-tors was evaluated using the Mann-Whitney test. The differences were considered significant at p<0.05.

R E S U L T S In the animals of the main group, we observed a

short-term period (during the first 3 days) of a decrease in the content of both glucorticoid (p<0.05) and mineralocorticoid (p<0.01) receptors (Fig.1, 2). This fact can be explained by the possible structural and functional rearrangement of physiological/biochemi-cal systems for the implementation of a complex of ad-aptation reactions to the changed internal conditions. Starting from the 5th day of the post-resuscitation period, we observed a gradual increase in the activity of glucocorticoid and reduced activity of mineralocor-ticoid receptors. Moreover, the level of glucocorticoid receptors gradually increased, and by the end of the follow-up period was significantly higher by 23% (p<0.05) (Fig. 1).

The dynamics of the increase in the level of mineralocorticoid receptors were recorded 2 weeks after ischemia-reperfusion injury (Fig. 2). By the end of the follow-up period, the level of mineralocorticoid receptors did not significantly differ from those in the control group of animals.

Fig. 1. Dynamics of the content of glucocorticoid receptors in the kidneys of rats in the post-resuscitation period (*p<0.05)

D I S C U S S I O N Adaptation to hypoxia is a reaction aimed at

maintaining the vital activity of the body in condi-tions of oxygen deficiency. It is controlled by central, intercellular, and intracellular regulatory mechanisms. During the period of an urgent generalized response to hypoxia, there is a simultaneous activation of various signaling systems of regulation and an increase in the urgent resistance of the body to oxygen deficiency [4]. The leading role in this process is played by the hy-pothalamus—pituitary—adrenal system and its main mediators-catecholamines and corticosteroids. During this period, multiple subordinate signaling systems are also activated, which ensure the formation of an urgent compensatory protective reaction of the body in response to hypoxia [1].

The kidney is one of the main target organs for systemic circulatory disorders. Studies of various scien-tists have shown that aldosterone, through mineralo-corticoid signaling, plays a key role in the pathogenesis of ischemia-reperfusion kidney damage, manifested in the development of progressive renal dysfunction, proteinuria, glomerulosclerosis, tubulointerstitial fibrosis, etc. [2, 3, 5]. According to Barba-Navarro R. I. blockage of mineralocorticoid receptors with spironolactone before or even after ischemia prevents acute functional and structural damage caused by ischemia-reperfusion [6].

C O N C L U S I O N After ischemia-reperfusion injury, a dynamic

increase in the level of glucocorticoid receptors in the kidney is observed from day 5. Recovery of the level of mineralocorticoid receptors is observed from the 14th day. Consequently, in the early post-resuscitation period, the kidney is most vulnerable to the effects of remodeling factors and fibrosis.

R E F E R E N C E S1. Bayburina G.A., Nurgaleeva E.A., Agletdi-

nov E.F., Samigullina A.F. Effect of hypoxia toler-ance on the relation between indicators of free radical oxidation of lipides and proteins in murine kidneys during the post-resuscitation period // Kazan medical journal. – 2017. – Vol. 98. – No 6. – P. 949–954. doi: 10.17750/KMJ2017-949

2. Jaisser F., Farman N. Emerging roles of the mineralocorticoid receptor in pathology: toward new paradigms in clinical pharmacology. Pharmacol. Rev. 2016; 68(1): 49–75. DOI: 10.1124/pr.115.011106

3. Zheng HF, Zhang YN, He JN, Yang Z, Zhang R, Li L, Luo ZH, Ye YR, Sun QQ. Hydroxychloro-quine Inhibits Macrophage Activation and Attenuates Renal Fibrosis After Ischemia-Reperfusion Injury. Frontiers in immunology. 2021;12: 645100. DOI: 10.3389/fimmu.2021.645100



Fig. 2. Dynamics of mineralocorti-coid receptor content in rat kidneys in the post-resuscitation period

4. dos Santos MS, Canale D, Bernardo DRD, Shimizu MHM, Seguro AC, Volpini RA, de Braganca AC. The Restoration of Vitamin D Levels Slows the Progression of Renal Ischemic Injury in Rats Previously Deficient in Vitamin D. Fron-tiers in medicine. 2021; 8: 625647. DOI: 10.3389/fmed.2021.625647

5. Blankenburg M., Fett A.-K., Eisenring S., Haas G., Gay A. Patient characteristics and initiation of mineralocorticoid receptor antagonists in patients with chronic kidney disease in routine clinical practice in the US: a retrospective cohort study. BMC Nephrol. 2019; 20(1): 171. doi: 10.1186/s12882-019-1348-4.

6. Barba-Navarro R., Tapia-Silva M., Garza-Garcia C., López-Giacoman S., Melgoza-To-ral I., Vázquez-Rangel A., Bazúa-Valenti S., Bobadilla N., Wasung de Lay M., Baranda F., Chawla L.S., Gamba G., Madero M. The effect of spironolactone on acute kidney injury after cardiac surgery: a randomized, placebo-controlled trial. Am. J. Kidney Dis. 2017; 69(2): 192–199. DOI: 10.1053/j.ajkd.2016.06.013.



BACTERIAL SYMBIOSIS IN COMPLICATED ULCERS: THE PATHOGENETIC HYPOTHESIS

A B S T R A C T — Aim of this paper is to estimate crystallogenic properties of gastric mucosa in connection with its microbial contamination. We investigated crystallogenic properties of some biological substrata (gastric mucosa, gastric mucosal layer homogenates) in 12 healthy individuals and 30 patients with ulcer disease complicated in 12 cases by perforation, bleeding or penetration. Biological substrata were received at fibrogastroduodenoscopy. Estimation of crystallogenic properties of biological material was accomplished by classic crystalloscopy. Biological substrata crystalloscopic investigation was accompanied by its traditional microbiological study for Helicobacter pylori and detection of other microorganisms. Our data allow to suppose dual contamination of stomach mucosa both by Helicobacter pylori and Providencia or Morganella. This combination caused elevation of gastric mucosa crystallogenic properties that provoked formation of ulcer. Procrystallogenic potential of this symbiosis may be an important link to the pathogenesis of ulcer disease which realized through microorganism-associated mucosal damage and the progression of complications.

K E Y W O R D S — ulcer disease, microbe-associated crystallogenesis, Helicobacter pylori, Providencia, Morganella.

Andrew Martusevich1 , Giulnar Orudzhova1 ,Anastasia Romanova2 , Oksana Shubina1

1 Privolzhsky Research Medical University, Nizhny Novgorod;2 Kirov State Medical University, Kirov, Russia

[email protected]


I N T R O D U C T I O NMicroogranism susceptibility to crystallization

was stated long ago. So, it was shown by V.F. Chubu-kov (1982), that bacteria can form many variants of crystal and pseudocrystal structures [1]. These data was confirmed by other investigators [2, 3, 5, 6]. We named illustrated phenomenon as «microorganism-associated crystallogenesis» (MAC). In our opinion, it has number of functions, such as protective, patho-genic etc [2, 3, 5]. Literature data analysis has shown, that in natural conditions a protective function of MAC is dominated, and a pathogenic function realizes at bacterial antagonism or infection process [2].

Special variant of MAC is the bacterial symbiosis with a high crystallogenic activity. Example of this sym-biosis is microbial induced film-formation in catheters.

Aim of this paper: to estimate crystallogenic properties of gastric mucosa in connection with its microbial contamination.

M A T E R I A L A N D M E T H O D SWe investigated crystallogenic properties of some

biological substrata (gastric mucosa, gastric mucosal layer homogenates) in 12 healthy individuals and 30 patients with ulcer disease, among them 12 patients with ulcer complicated by perforation, bleeding or penetration. Biological substrata were obtained using fibrogastroduodenoscopy.

Estimation of crystallogenic and initiated proper-ties of biological material was accomplished by own methods (classic crystalloscopy and comparative tezi-ography [4]). We used 0,9% sodium chloride solution as basic substance in teziographic test.

Biological substrata crystalloscopic investigation was accompanied by its traditional microbiologi-cal study for Helicobacter pylori (Нр) or/and other microorganisms detection.

Statistical processing of the results was performed using variation statistics algorithms using Microsoft Excel 2007 and Statistica 6.1 for Windows.

R E S U L T SInvestigation of own and initiated crystallogenesis

of gastric mucosa in individuals without gastrointesti-nal pathology allows to state that in this case biological material has a low crystallogenic activity. At dehydra-tion it mainly formed numerous amorphic structures and single small crystals. In described crystalloscopic picture of gastric mucosa Helicobacter pylori was not detected in biological substrate. So, in physiological conditions bacterial flora and gastric mucosa con-tain bulk mucopolysaccharides [3], have no essential crystallogenic potential and do not promote material initiation of basic substance crystallogenesis in tezio-graphic test (Fig. 1).

On endoscopy, the evidence of gastric ulcer induced by Hp infection, crystallizability and initia-tion potential of gastric mucosa are meaningfully higher than in healthy people (p<0,05). It manifested as elevation of the portion of crystalloid structures in facia caused by crystallizability rate augmenta-tion. However, this tendency is not accompanied by complication of structural elements (structure index saving at normal level; p>0,05). These changes were


http://dx.doi.org/10.35630/2199-885X/2021/11/2/7

https://orcid.org/0000-0002-0818-5316

https://orcid.org/0000-0003-0447-5970

https://orcid.org/0000-0003-2219-2835

http://dx.doi.org/10.35630/2199-885X/2021/11/2/7


corroborated by morphological and morphometric analysis of gastric mucus layers from homogenate mu-cosal samples. In their facias only single small crystals are presented.

At complicated variants of ulcer disease notice-able activation of crystallogenesis in investigated biological fluids is visualized. So, in this case crys-tallograms for homogenates of gastric mucosa and gastric mucosal layer include numerous single-crystal elements and dendrites. It caused elevation of crys-tallizability and structure index level in comparison with healthy people and patients with non-compli-cated ulcer disease rate. It is interesting, that form-ing crystals have rather low degree of destruction (1,42±0,31 rel. un.), which can indicate its importance in this pathogenic type of ulcer disease. This tendency is visualized in teziographic facias too. At morphologi-cal analysis in homogenates of the gastric mucosal layer, its high crystallogenic properties were discov-ered. Crystallograms of this biological substrate con-tained dendrites. It is very important, that at microbio-logical study in all biological materials Providencia or Morganella bacteria were marked out in addition to Нр. Based on the above consideration we assume that its symbiosis can be an initial factor of the damaged gastric mucosa.

C O N C L U S I O NOur data allow supposing dual contamination

of gastric mucosa both by Helicobacter pylori and Providencia or Morganella. This combination caused elevation of gastric mucosa crystallogenic properties that provoked formation of ulcer. Procrystallogenic potential of this symbiosis may be an important link

Fig. 1. Visuometric estimation of crystallo-genic properties of gastric mucosa (Cr — crystallizability, SI — structure index, FDD — facia destruction degree, CUD — complicated ulcer disease)

to ulcer development, which realized through micro-organism-associated mucosa damage (result of MAC activation) and progression of the complications.

R E F E R E N C E S1. Chubukov V.F. Microbes store metals // Chemistry

and life. – 1982. – Vol. 11. – P. 53–55.2. Dieudonné A., Pignol D., Prévéral S. Mag-

netosomes: biogenic iron nanoparticles produced by environmental bacteria // Appl Microbiol Biotechnol. – 2019 Vol. 103, N9. – P. 3637–3649. doi: 10.1007/s00253-019-09728-9.

3. Faivre D., Godec T.U. From bacteria to mollusks: the principles underlying the biomineralization of iron oxide materials // Angew Chem Int Ed Engl. – 2015. – Vol. 54, N 16. – P. 4728–4747. doi: 10.1002/anie.201408900.

4. Martusevich A.K., Kamakin N.F. Unified algorithm for the study of free and initiated crystal-logenesis of biological fluids // Clinical laboratory diagnostics. – 2007. – No. 6. – P. 21–24.

5. McCausland HC, Komeili A. Magnetic genes: Studying the genetics of biomineralization in magne-totactic bacteria // PLoS Genet. – 2020. – Vol. 16, N 2. – e1008499. doi: 10.1371/journal.pgen.1008499.

6. Zhang K, Xue Y, Zhang J, Hu X. Removal of lead from acidic wastewater by bio-mineralized bacteria with pH self-regulation // Chemosphere. – 2020. – Vol. 241. – 125041. doi: 10.1016/j.chemos-phere.2019.125041.



STUDY OF POSSIBLE MECHANISMS OF ACTOPROTECTIVE ACTION OF СATEKHIN HYDRATE

A B S T R A C T — A study was carried out to study the possible mechanisms of actoprotective action of catechin hydrate administered at a dosage of 100 mg/kg. The animals were subjected to daily exhausting loads in the forced swimming test with a load of 10% of the animal's weight. After the end of the experiment, the rats were decapitated under chloral hydrate anesthesia (350 mg/kg) and the skeletal muscle was taken to obtain the supernatant. Using the ELISA method, the concentrations of nitric oxide isoforms (eNOS, iNOS, nNOS), PPAR (peroxisome proliferator-activated receptors) and JNK were estimated.It was found that against the background of the introduction of the test substance, an increase in eNOS activity was observed by 48.7% (p <0.05) relative to the group of negative control rats, as well as a decrease in iNOS and JNK — 1.9 times (p <0.05). In comparison with the group receiving Metaprot®, the concentration of endothelial synthase in the group receiving catechin hydrate was 1.3 times higher (p <0.05).The experiment performed suggests that the actoprotective effect of catechin hydrate is likely to be associated with inhibition of pathways mediated by JNK, activation of PPAR receptors, and the effect of these compounds on NO isoforms.

K E Y W O R D S — actoprotectors, mechanism of action, rats, catechin hydrate.

Anastasia Gerashchenko1 , Natalia Shabanova1 ,Marina Efremova1, Andrey Voronkov2

1 Pyatigorsk Medical and Pharmaceutical Institute — Branch of Volgograd State Medical University, Pyatigorsk;2 The Volgograd State Medical University, Volgograd, Russia

[email protected]

Received 03 March 2021 Received in revised form 17 April 2021; Accepted 25 April 2021

are often exposed to, have an effect on all systems of the body, including skeletal muscles. It is known that the symptomatology of musculoskeletal fatigue combines factors that limit the activity of skeletal muscles, first of all: a decrease in the level of blood flow in skeletal muscles and the intensity of metabolic processes in muscle tissue. In this case, the leading role in maintain-ing the proper level of muscle blood flow is assigned to the vascular endothelium [4]. It has been established that endothelial dysfunction developing during physi-cal exertion is associated with a decrease in the catalytic activity of endothelial nitric oxide synthase (eNOS), one of the key endothelial enzymes that provides the physiological secretion of nitric oxide (NO). In ad-dition, it is worth noting that optimal blood flow in skeletal muscle is influenced by compounds that affect PPARs (Peroxisome Proliferator Activated). This type of receptor belongs to the superfamil nuclear glyco-proteins activated by ligand transcription factors, of synthetic or endogenous origin.

Along with oxidative stress, apoptosis is also one of the main pathogenetic mechanisms of destruction of skeletal muscle myocytes under conditions of intense muscle work, and its modulation is a new promising direction for restoring the functional activity of striated muscles. According to recent studies, c-Jun-terminal kinase ( JNK) may be a potential target for regulating apoptotic processes [5].

Thus, based on the available literature data, it can be assumed that by acting on specific pathological targets, it will be possible to reduce the manifestations of muscle dysfunction.

Objective: to study the possible mechanisms of the actoprotective action of catechin hydrate.

M A T E R I A L S A N D M E T H O D SThe experiment was carried out in accordance

with the "Guidelines for the conduct of preclinical studies of drugs, ed. A.N.Mironov (2012 ed.) [6]. The animals were kept in the vivarium of the PMFI-branch of the Volgograd State Medical University (Volgograd, Russia). The study was carried out on male Wistar rats weighing 220–230 g. All animals were divided into 4 experimental groups. During the experiment, the rats

I N T R O D U C T I O NCatechin is flavan-3-ol, a flavan derivative with

four phenolic hydroxyl groups, which exhibits a wide range of pharmacological activities, including: antioxi-dant, antihypoxic, antibacterial [1, 2]. It is also worth noting that this compound is capable of increasing physical performance against the background of exhausting physical exertion [3]. In view of its poly-functional action, this compound can be classified as a group of actoprotectors.

Physical activity in natural conditions, and some-times even exhausting physical activity, which athletes


http://dx.doi.org/10.35630/2199-885X/2021/11/2/8

https://orcid.org/0000-0003-0294-2926

https://orcid.org/0000-0002-7693-5182

http://dx.doi.org/10.35630/2199-885X/2021/11/2/8


were kept in standard vivarium conditions (natural mode of light change, temperature, relative humidity, standard diet of laboratory animals, weekly change of bedding and cages, fixed time of feeding and drinking) in compliance with the International Recommenda-tions of the European Convention for the Protection of Vertebrate Animals used in experimental studies. The animals were subjected to daily exhausting loads in the Forced Swimming test with a load of 10% of the animal's weight [6]. Positive control (PC), the first group of animals (rats swam with days of rest), the second — negative control (NC) group of untreated animals (0.9% sodium chloride solution in an equiva-lent volume), the third — received the studied sub-stance catechin hydrate at a dosage of 100 mg/kg [7], the fourth group received actoprotectorMetaprot® at a dosage of 25 mg/kg [8].

The substance catechin hydrate, as well as the reference drug, were administered per os for 10 days 60 minutes before the expected load. After the end of the experiment, the animals were decapitated under chloral hydrate anesthesia (350 mg/kg) and the skeletal muscle was taken to obtain the supernatant. In the obtained biomaterial, using the ELISA method on a microplate reader (Tecan Infinite F50, Austria), the concentrations of nitric oxide isoforms (eNOS, iNOS, nNOS), PPAR (peroxisome proliferator-activated receptors) and JNK (c-Jun-terminal kinase) were estimated. The results were processed using the STATISTICA 6.0 software.

R E S U L T SDuring the experiment, it was found that in the

group of rats without pharmacological support (NC), after prolonged depleting loads, there was a decrease in the concentration of (eNOS) — endothelial synthase relative to the PK group by 53% (p <0,05), while the concentration of inducible NOS was 2,7 times higher in comparison with the same group (p <0,05). Relative to the PK group, a significant increase in the concen-tration of PPAR and JNK was found 4,6 and 5,1 times (Fig. 1).

The course use of catechin hydrate led to an increase in the concentration of eNOS — by 48,7% (p <0,05) and a decrease in iNOS, as well as JNK — by 1,9 times (p <0,05) (Fig. 2). However, it should be noted that the concentration of endothelial NOS was 1,3 times higher than that of the group receiving Met-aprot® at a dosage of 25 mg/kg (p <0,05). The PPAR concentration index did not differ statistically signifi-cantly from the group of NС rats.

The introduction of Metaprot® promoted an increase in PPAR concentration by 2,7 times (p <0,05) and a decrease in JNK by 1,7 times (p <0,05) in rela-tion to the group of NK animals. Against the back-

Fig. 1. Changes in the concentration of PPAR and JNK against the background of the introduction of the studied substance catechin hydrate in the supernatant of the skeletal muscle of rats exposed to exhausting physical exertion

Note: μ — statistically significant relative to the group of animals PK (Student's t-test, p <0,05); * — statistically significant relative to the group of NC animals (Student's t-test, p <0,05)

Fig. 2. Changes in the concentration of NOS isoforms against the background of the introduction of the studied substance catechin hydrate in the supernatant of the skeletal muscle of rats subjected to exhausting physical exertion

Note: μ — statistically significant relative to the group of animals PK (Student's t-test, p <0.05); * — statistically significant relative to the group of NC animals (Student's t-test, p <0.05); — statistically significant relative to the group receiving Metaprot®

ground of administration of the drug Metaprot®, there was no significant effect on the content of nitrogen monoxide isoenzymes in comparison with the group of NK rats.

It should be noted that neither the use of catechin hydrate nor Metaprot® led to a significant change in the expression of the isoenzyme of neuronal synthase (nNOS).



С O N C L U S I O NThe use of catechin hydrate increased the catalytic

activity of eNOS by 48,7% (p <0,05) and 1,3 times (p <0,05), relative to the group of NK rats and the group receiving Metaprot, respectively. Against the back-ground of oral intake of the test substance, a decrease in the concentration of JNK of 64,3% (p <0,05) was noted relative to the group of NK animals. As a result of the experiment, it can be assumed that the acto-protective effect of catechin hydrate may possibly be associated with inhibition of pathways mediated by JNK, activation of PPAR receptors, and the effect of these compounds on NO isoforms.

R E F E R E N C E S1. Yang M.J., Hung Y.A., Wong T.W., et al. Effects

of Blue-Light-Induced Free Radical Formation from Catechin Hydrate on the Inactivation ofAcinetobac-terbaumannii, Including a Carbapenem-Resistant Strain // Molecules. – 2018. – vol.23. – P.1631. doi.org/10.3390/molecules23071631

2. Hi M., Nie Y., Zheng X.Q., et al. (UVB) photo-sensitivities of tea catechins and the relevant chemical conversions // Molecules. – 2016. – vol.21. – P. 1345.doi.org/10.3390/molecules21101345

3. Voronkov A. V., Oganesyan E. T., Gerash-chenko A. D. Aspects of actoprotective activity of

some natural compounds of different chemical struc-ture // Sports medicine: science and practice. – 2017. – T. 7. – No 1. – P. 92–96.(In Russ.)

4. Eitenmuller I., Volger O., Kluge A. [et al.]. The range of adaptation by collateral ves-sels after femoral artery occlusion // Circ Res. – 2006. – vol. 99. – P. 656–662. doi.org/10.1161/01.RES.0000242560.77512.dd

5. Mironov A.N. The guidelines for preclinical studies of pharmaceuticals. Part one. – M.: Grif and K, 2012. – 944 p. (In Russ.)

6. Porsolt R.D., Anton G., Blavet N., Jalfre М. Behavioral despairin rats: anewmodel sensitive to antidepressant treatment // Europ. J. Pharmacol.- 1978. – vol. 47. – Р. 379–391. doi.org/10.1016/0014-2999(78)90118-8

7. Voronkov A.V., Abaev V.T., Oganesyan E.T., Pozdnyakov D.I. Study of the influence of ATACL substance on the physical and mental state of animals under conditions of prolonged exhausting loads // Modern problems of science and education. – 2015. – No 3. – P. 628. (In Russ.).

8. Gerashchenko A.D, Pozdnyakov D.I. Natural polyphenolic compounds in the pharmacological correction of extreme conditions // Pharmacy and Pharmacology. – 2015. – Vol. 3. – Р. 35–36.



PHARMACOKINETICS AND PHARMACODYNAMICS OF CHAGA BIRCH MUSHROOM COMPONENTS (INONOTUS OBLIQUUS)

A B S T R A C T — The study presents an analysis of the available scientific literature and outcomes of clinical trials on biologically active components of the birch Chaga mushroom Inonotus obliquus (I. obliquus). Our experimental and clinical studies were aimed at testing a novel plant-based substance for patients with cancer or tumors of various locations. The paper investigates pathogenetic mechanisms and effectiveness of chaga mushroom extracts depending on the topography of the tumor, the stage of the disease and the age of patients, as well as the questions of pharmacokinetics and pharmacodynamics of the components of I. obliquus. As a result of the analysis of the obtained data, the pathogenetically justified clinical application of extracts of the birch chaga mushroom I. obliquus in patients with oncological pathology was conceptually presented. The study allowed us to reveal possible mechanisms of pathogenetic effects of the birch chaga mushroom (Inonotus obliquus) extracts obtained by different extraction methods. Analysis of the available data enabled us to deepen our understanding of the effectiveness and mechanisms of the effect of the I. obliquus extracts on tumors of various localization. Finally we present recommendations on predicting the possibility of using the I. obliquus to improve outcomes in patients with cancer of various etiology and locations.

K E Y W O R D S — chaga mushroom, Inonotus obliquus, inotodiol, glycans, neuroprotector, hyperglycemia, antioxidants, stroke, apoptosis, proliferation, regeneration, malignancy, cancer.

Pavel Khoroshutin1, Galina Reva2,3 ,Tatsuo Yamamoto3, Ivan Reva3

1 Baikal Herbs Ltd., Science and quality management department laboratory, Irkutsk 2 Far Eastern Federal University, Vladivostok, Russia3 International Medical Education and Research Center, Niigata, Japan, Vladivostok, Russia

[email protected]

Received 11 May 2021; Received in revised form 28 May 2021; Accepted 1 June 2021

R E L E V A N C EAt the current stage of development of medicine

it is of particular importance to search for substances that can be used in prevention and supporting meas-ures during treatment of cancer and neurodegenerative diseases. In contrast to strong synthetic drugs with nu-merous negative side effects which sometimes exceed

their benefit, medicinal plants can be assumed to be effective and safe. The analysis by Yen-Nien Hou, Gary Deng, Jun J. Mao (2020) of the data from the Integra-tive Medicine Service at Memorial Sloan Kettering Nassau cancer center (USA) allowed us to establish that chaga mushroom is in the first place out of the 10 most popular herbs/supplements in 2018 [11, 33]. The chaga mushroom, Inonotus obliquus (I. obliquus), has been a well-known traditional medicine remedy since the time of Avicenna [1, 3]. However, the systematic pharmacological study of the active substances of I. obliquus was started only in the middle of the 20th century, when fundamental medicine claimed the medicinal advantages of the fungus Inonotus obliquus in experimental studies with the suppression of cancer cell growth [26]. The chemical composition and spec-trum of the therapeutic use of Inonotus obliquus as raw material, as well as methods for assessing the resource potential of this fungus, are presented in numerous studies despite the background of insufficient data on the molecular mechanisms of action of mushroom extracts [17]. At the present stage, the medicinal prop-erties of I. obliquus have been studied only by 10% [28]. At the same time, the available scientific literature on this issue considers to a greater extent the results of experiments performed on mice, mongrel and geneti-cally modified, or on Danio fish, which is a key factor for choosing the direction of research in the study and systematization of data on the mechanisms of their action on the human body.

Over the past fifty years, ideas about the use of chaga extracts as additives to essential medicines are based mainly on the results of preclinical studies, with a small number of adequately planned experiments and a limited number of studies conducted on cancer patients with cancer of various topography and etiol-ogy [4]. Therefore, despite numerous experimental and biochemical studies, the available data on the biologi-cal effects of individual components at the present stage does not allow us to give an exhaustive pathoge-netic justification for the use of popular additives from chaga mushroom, registered and obtained by various methods [24]. There is also insufficient information to fully decipher the mechanisms of action, substanti-ate the effectiveness of drugs from the chaga mush-room, prove their advantages and minimize the risks


http://dx.doi.org/10.35630/2199-885X/2021/11/2/9

http://dx.doi.org/10.35630/2199-885X/2021/11/2/9


of side effects. This analysis of available data is also not presented in the archive database of the Integra-tive Medicine Service at Memorial Sloan Kettering cancer center, which provides a summary of research results indicating the intended uses, side effects, and interactions of herbal medicines for approximately 284 dietary supplements [10].

The known antitumor effect of glycans and glucans that are part of I. obliquus is mainly associ-ated with the induction of increased immune activity, and their effect in the composition of chaga extract on malignant tissue has not been sufficiently studied, there are only a few experiments in Vitro confirming their inhibitory effect on the proliferation of cancer cells [5].

Critical unsolved problems in the use of chaga mushroom (I. obliquus) extracts and the lack of systematic analysis of available materials on this issue require scientific research to be directed on these mat-ters.

The aim of the study was to generalize data on the medicinal properties of the birch chaga mushroom (I. obliquus) and conduct a comparative analysis of the results of its usage in tumors of various locations and etiology.

In this regard, the following tasks were solved:1. To provide an analysis of available literature

data about the current state of the studied issue.2. To obtain a comparative description of the

results of treatment of cancer patients with tumors of different localization and etiology.

3. To determine the possible mechanisms of pathogenetic effects of extracts of the birch chaga mushroom (Inonotus obliquus) obtained by different extraction methods based on the available data.

4. To evaluate the effectiveness of the extracts of Inonotus obliquus on tumors of various localization.

5. Conceptually substantiate the clinical use of Inonotus obliquus extracts in patients with cancer.

M A T E R I A L S A N D M E T H O D SWe reviewed available literature data on clinical

use of extracts of the Inonotus obliquus for the last 20 years, as well as available archival and scientific materi-als on the results of treatment of patients with cancer of various etiologies and topography of maligning structures.

R E S E A R C H R E S U L T S In contrast to the studies by Draggendorf G.L.

(1864) [7], which did not discover useful substances in the birch chaga mushroom (Inonotus obliquus) and the presence of alkaloids and glycosides in its content, many researchers found more than 10 amino acids,

40% of which are glutamic acid, tyrosine, serine, threonine, leucine, methionine, histidine and lysine, as well as polysaccharides, in addition to numerous other chemical elements [6, 19]. Previously, data on the health properties of fungal polysaccharides were obtained by analyzing the chemical composition of 651 species and 7 infraspecific taxa from 182 genera of higher homo- and heterobasidiomycetes. It was found that, despite the different chemical composition, the majority of fungal polysaccharides belong to the group of beta-glucans, they have beta (1-->3) bonds in the main chain of glucan and additional beta (1-->>6) branching points that cause their antitumor effect. Most of the clinical evidence for antitumor activity comes from the polysaccharides like lentinan, PSK (krestin), and schizophyllan. It was found that the ef-fectiveness of high-molecular-weight glucans over-whelms than that of low-molecular-weight glucans.

Satoru Arata, Jun Watanabe, and Masako Maeda (2016) studied the actual effect and underlying mecha-nisms of the effect of continuous intake of an aqueous extract from I. obliquus on the suppression of Lewis lung carcinoma growth and spontaneous metastasis in mouse models [2]. After 3 weeks of continuous intake of the extract at a dose of 6 mg/kg/day, the authors obtained significant suppressive effects: in models of tumor-bearing mice, a 60% reduction in the tumor was observed, while in metastatic mice, the number of maligning nodules decreased by 25% compared to the control group. In addition, mice treated with I. obliquus extract showed not only tumor reduction, but also inhibition of vascularization.

Eid J.I., Das B. (2020) consider Inonotus obliquus to be one of the most powerful antioxidants in the world [8]. The authors note that the therapeutic effects of chaga components are well characterized only in vit-ro; the effects of development in vivo are not described in detail. Using the example of Danio fish, the authors identified the influence of I. obliquus on the cell cycle and apoptosis. Polysaccharides of chaga mushrooms, in addition to phenolic compounds, included xylulose, rhamnose, mannose, glucose, inositol, and galactose. Staining with the DNA-binding dye acridine orange showed that polysaccharides of Inonotus obliquus allevi-ate oxidative stress [8]. Flow cytometric analysis using H2DCFDA, which specifically binds to fragmented DNA in cells, showed significantly reduced levels of intracellular reactive oxygen species (ROS) (p <0.05), which in turn reduced apoptosis in developing em-bryos. Analysis of the cell cycle by measuring the DNA content using flow cytometry showed that chaga polysaccharides moderately delay cells at the G1 stage, thereby inhibiting cell proliferation, which can be fur-ther studied in cancer research. In general, temporary



exposure to chaga polysaccharide extract reduced the volume of intracellular reactive oxygen species (ROS) and contributed to the normal development of Danio fish.

Li Z., Mei J., Jiang L., Geng C., Li Q., Yao X., Cao J. (2019) found that tacrine, the first drug licensed for the treatment of Alzheimer's disease, induces apop-tosis in HepG2 cells by generating reactive oxygen species (ROS) and mitochondrial dysfunction [18]. The authors studied the possible protective effect of polysaccharides from I. obliquus, obtaining evidence that I. obliquus polysaccharides reduce tacrine-induced apoptosis in HepG2 cells. The experiment also observed inhibition of tacrine-induced ROS genera-tion, 8-OHdG formation in mitochondrial DNA, and loss of mitochondrial transmembrane potential when using I. obliquus polysaccharides. In addition, the intake of I. obliquus extract reduced the release of cytochrome C and activation of caspase-3 induced by tacrine. These data suggest that fungal polysaccharides may inhibit tacrine-induced apoptosis in HepG2 cells. This protection is mediated by an antioxidant cyto-protective mechanism [18]. Similar hepatoprotective properties of I. obliquus extract were proved by studies of Hong K.B., Noh D.O., Park Y., Suh H.J. (2015) [10].

Nguyen T.M.N., Le H.S., Le B.V. et al. (2020) note that the lanostane triterpenoid, inotodiol, is found exclusively in the chaga mushroom; the authors found evidence that purified inotodiol has activity to suppress mast cell function in vivo [23]. Inotodiol also relieved symptoms similar to those observed in the ovalbumin-induced (cOVA) mouse food allergy model. As with untreated 70% ethanol extract of chaga mushroom (320 mg/kg), oral administration of inotodiol (20 mg/kg), both for prevention and treatment, was accompanied by a significant reduc-tion in the manifestations of allergic symptoms and inflammatory lesions in the small intestine that appear after repeated oral administration of cOVA. Despite the fact that inotodiol (20 mg/kg) and chaga mush-room extract (320 mg/kg) acted to the same extent, it was found that the immunological mechanisms underlying these effects differ from each other. The authors illustrated the evidence obtained from several in vivo analyses, including experiments with passive systemic anaphylaxis mediated by mast cells, activation and proliferation of adaptively transferred antigen-specific T cells, and production of IgG1, IgE, and IgA immunoglobulins by antigen-specific B cells. Inoto-diol selectively inhibited mast cell function without significantly affecting other immune responses, while crude chaga mushroom extract randomly suppressed a variety of immune responses. This indicates the presence of several immunomodulating biologically

active components in the chaga mushroom that act on various targets. The strong anti-allergic activity of inotodiol, as well as its remarkable selectivity against mast cells, makes it an excellent tool of choice for the treatment of food allergies with high efficiency and no side effects.

Many authors have described the positive anti-tumor effect of I. Obliquus and its extracts containing the trioterpenoid inotodiol. Nikitina S.A., Khabibra-khmanova V.R., Sysoeva M.A. (2016) summarized data on the chemical composition of triterpene and steroid compounds isolated from chaga mushroom grown in natural conditions or in synthetic culture [25]. Particular attention was paid to the biological activity of chaga mushroom extracts and these specific compounds against various cancer cell lines in vitro and in vivo. This analysis demonstrated some common features in the inhibition of growth of various cell lines by components of the chaga fungus. In this context, triterpene compounds containing an OH group at C22 and a side chain unsaturated bond are the most active [25]. Zhang P., Cao X., Li C., et al. (2016) stud-ied the role of the squalene synthase gene isolated from Inonotus obliquus in triterpene synthesis by detecting squalene in vitro in a reaction mixture of particles us-ing high-performance liquid chromatographic analysis [34]. The positive effect of treatment with the use of triterpenoid inotodiol have been demonstrated in hepatoma, leukemia, colon cancer and cervical cancer. Anti-carcinogenic effects of cell growth inhibition in lung carcinoma have also been shown. Due to the identified oncostatic activity of I. obliquus, Sun Y., Yin T., Chen X. H., Zhang G., et al. (2011) suggested using polysaccharide-triterpenoid complexes of I. Obliquus as components of heterogeneous inhibitors of cancer cell proliferation [28]. Zhang S.D., Yu L., Wang P. Kou, Li J. et al. (2019) studied the potential mechanisms of inotodiol for HeLa cell migration, invasion and apoptosis via the p53-dependent pathway, transwell invasion, flow cytometry, caspase-3 activity analysis and Western blot analysis, as well as the participa-tion of the p53 signaling pathway in anti-metastatic and Pro-apoptosis [35]. In addition, the function of the p53 tumor suppressor was further verified by small interfering RNA. The dependence of the effect of inotodiol on its concentration was established. The cytotoxic effect on HeLa cells at concentrations above 25 microns significantly inhibits the growth of HeLa cells and even induces apoptosis. This result was further confirmed by analysis of cell proliferation and morphology. Analysis of in vitro wound healing and transwell invasion showed that treatment with inoto-diol in low concentrations significantly inhibits cell migration and invasion in a dose-dependent manner.



Reduced levels of matrix metallopeptidase-2 (MMP2) and matrix metallopeptidase-9 (MMP9) also depend on the drug concentration. Inotodiol significantly induces apoptosis of tumor cells through its effect on annexin-V-FITC, which is associated with activation of pro-apoptotic PARP proteins, partition of caspase-3 and Bax expression, and inhibition of the anti-apop-totic Bcl-2 protein expression. The antitumor activity of inotodiol can be inhibited by switching off the tumor suppressor p53. Pretreatment of p53-specific small interfering RNA (si-p53) significantly inhibits inotodiol-induced apoptosis of HeLa cells and reduces the activity of caspase-3. Moreover, the inhibitory effect of inotodiol on tumor migration and invasion was blocked by p53 knockdown.

Zhang X., Bao C., Zhang J. (2018) investigated the effect of inotodiol (lanostane triterpenoid) on the cellular growth of breast tumors in diabetic condi-tions [36]. The research was conducted on a model of breast cancer against the background of diabetes in female Sprague-Dowley rats, caused by the ad-ministration of streptozotocin (STZ) at a dose of 35 mg/kg, followed by the induction of breast cancer 7,12-dimethylbenzanthracene (DMBA) at a dose of 10 mg/kg. Histological examination of the pancreas and of breast tumor proliferation was conducted by immu-nohistochemical methods with PCNA staining, Tunel method for apoptosis. The results of the study showed that inotodiol reduces blood glucose levels in SD rats, as well as reduces the level of cholesterol, triglycerides and high-density lipoproteins in blood plasma. Expres-sion of the PCNA proliferation marker was reduced when taking inotodiol. It lowered the expression of β-catenin and its downstream targets (c-Myc and cyclin D1) with subsequent induction of apoptosis. Evidence-based results indicate that inotodiol regu-lates blood glucose levels in diabetic rats, participates in regulating of proliferation while inhibiting breast tu-mor progression, inducing apoptosis through reduced regulation of β-catenin signaling [36].

Anti-carcinogenic effect with targeted effects of I. obliquus extracts are shown against several cancer cell lines in the absence of cytotoxic effects to normal cells. The optimal concentration of these complexes was estimated at 150 mg/ml. Results of oral doses of I. obliquus polysaccharide complexes showed suppres-sion of melanoma cell growth. Antihyperglycemic and antioxidant effects were demonstrated in experiments on mice with alloxan diabetes; data were obtained on lowering blood cholesterol levels and pancreatic regeneration. A number of studies have demonstrated the antimicrobial activity of unknown components of I. obliquus, which also have a clear bactericidal effect on a number of strains of Mycobacterium smegmatis and

Francisella tularensis. Based on these properties, dental films based on nano-particles of chaga have been developed for the treatment of inflammatory bacterial diseases of the oral mucosa.

Chaga mushroom glycans are considered patho-genic associated molecular patterns (PAMP), similar to molecules that bind to specific pattern recognition receptors (PRRs) on the surface of immune cells. This interaction induces activation of a signaling cascade that directs the synthesis of a specific repertory of im-mune effector molecules [30]. Wasser S.P. (2011, 2014, 2017) noted the importance of studying these natu-ral sources as a means of treating cancer [31]. Many fungal biopolymers have immunotherapeutic prop-erties, contributing to the inhibition of growth and inducing the destruction of tumor cells. Although the mechanism of their antitumor action is still not fully understood, the development of methods for stimulat-ing and modulating key immune responses with fungal polysaccharides is a central task in medicine in general and in oncology in particular [12]. Some of the fungal polysaccharides have passed through phase I, II, and III clinical trials and are widely and successfully used in Asia for the treatment of various types of cancer and other diseases [26]. Based on the chemical composi-tion, it is assumed that the components of the chaga mushroom have 126 therapeutic functions, including antitumor, immunomodulatory, antioxidant, anti-ischemic, cardiovascular, antihypercholesterolemic, antiviral, antibacterial, antiparasitic, antifungal, detoxifying, hepatoprotective and antidiabetic effects [32]. Given that the mechanism of their antitumor action has not yet been fully studied, the issues of stimulation and modulation of immune responses by chemical components of chaga are of key relevance at the present stage [29].

The results of more than 600 clinical studies testing the effects of fungi on humans have been published in numerous papers [21]. Mushrooms are an extensive and at the same time almost inexhaust-ible source of powerful new pharmaceutical products [25]. In particular, and this is the most important thing for modern medicine, they are an unlimited source of polysaccharides [4]. These polysaccharides have different chemical compositions, and most of them belong to the group of beta-glucans; they have beta (1-->3) bonds in the main chain of glucan and ad-ditional beta (1-->>6) branching points that provide mechanisms for their antitumor action. It appears that high-molecular-weight glucans are more effective than low-molecular-weight glucans [6]. The same opinion is shared by Xin X., Qu J., Veeraraghavan V.P., (2019), who showed existing achievements in research on the mechanisms of isolated fungal polysaccharides,



in particular (1-->3) -β-D-glucans, which can cause various cellular responses, such as the expression of cytokines and nitric oxide, which carry out antitumor mechanisms indirectly through stimulating T cells or binding other molecules of cellular immunity, such as polypeptides and proteins, conjugation of which always has a strong antitumor activity [32].

Cytotoxic bioactivity of I. obliquus was estab-lished in four human lung adenocarcinoma cell lines with different p53 status (A549, H1264, H1299, and Calu-6) [22]. According to the authors, the analysis of apoptosis induction tests with MeOH I. obliquus extract showed a decrease in cell viability in all lung cancer cell lines, and the cytotoxicity of these com-pounds was mediated by apoptosis through activation of caspase-3. Directed increase in bioactivity through fractionation of MeOH extract and chemical study of its cytotoxic hexano-soluble and Ch2Cl2-soluble frac-tions led to the isolation of eight triterpenoids, includ-ing a new lanostane-type triterpenoid called chagabu-sone A. The structures of the isolates were elucidated by high-resolution spectroscopic analysis. Among the isolated compounds, some of them showed the most powerful cytotoxic activity in all studied human lung cancer cell lines, with IC50 values varying from 75.1 to 227.4 microns.

In the treatment of cancer, fungi can inhibit metastasis due to the presence of arctigenin, lignan, in its composition—an important component and a representative of the group of phenolic compounds of plant origin. As is known, the basis of tissue malignan-cy inhibitors, cytostatics, contains organic compounds of the aromatic series, in whose molecules the hydroxyl groups OH− are bound to the carbon atoms of the aromatic ring. Arctigenin refers to phytoestrogens that are metabolized in the same way as estrogen hormones. The use is accompanied by a reduction in the risk of breast cancer after menopause, which is due to the fact that lignans have a milder effect on cells than human estrogen, effectively blocking the action of powerful endogenous human estrogen [27]. At the same time, the probability of malignancy in the mammary gland, which depends on growth hormones, is reduced by inhibiting individual proteins (NPAT proteins) neces-sary for the formation of cancer cells, paralyzing the ability of maligning cells to reproduce. Studies have shown that arctigenin is effective in the treatment of lung, liver, and stomach cancer [19].

Kang J.H., Jang J.E., Mishra S.K., et al. (2015) studied the effect of various fractions and components of the chaga mushroom (Inonotus Obliquus) on viabil-ity and apoptosis in human colorectal cancer (CRC) cell lines, and revealed the most effective inhibition of tumor growth by a single component identified

by pulsed nuclear magnetic resonance (NMR) as ergosterol peroxide with known antiproliferative and apoptotic activity [13]. Lee H.S., Kim E.J., Kim S.H. (2015) cultured human colon cancer HT-29 cells in the presence of 2.5–10 micrograms/ml of I. obliquus ethanol extract (IOEE) and analyzed cell cycle arrest by flow cytometry and Western blot protein expres-sion control. It was found that the treatment of cells with 2.5–10 micrograms/ml of IOEE reduces the number of viable HT-29 cells and DNA synthesis, increases the percentage of cells in the G1 phase, re-duces the expression of CDK2, CDK4 and cyclin D1 proteins, increases the expression of p21, p27 and p53, and inhibits phosphorylation of Rb and E2F1 expres-sion. Among I. obliquus fractions, fraction 2 (fraction-ated with dichloromethane) showed the same effect as EEIO treatment on cell proliferation and the level of protein associated with the cell cycle [16].

Currently, I. obliquus is considered a non-specific medicine for gastritis, gastric ulcer, polyposis and is used for precancerous therapy in liquid or tablet form, as a complex drug "Befungin" represented by a concen-trated extract of chaga mushrooms. To establish the pharmacological properties of I. obliquus, studies were conducted showing the anti-ulcer activity of ethanol extract of chaga. The anti-ulcer activity of I. obliquus was determined in rats with gastric ulcer (ethanol-induced ulcer). Ethanol extract of I. obliquus (200 mg/kg) did not cause any signs of toxicity or sensitivity in rats when administered orally. Oral administration of ethanol extract of I. obliquus showed anti-ulcer activity in all models used. The ethanol extract of I. obliquus showed effective anti-ulcer activity, which could be due to the presence of various biologically active compounds.

Na H.G, Park Y., Kim M.A., et al. (2019) noted that the use of chaga has numerous health benefits in alternative medicine in the treatment of obesity. Their proposed powder mixture of Chaga mushroom extracts was fermented using Lactobacillus acidophilus KCTC3925 (FCC), with the determination of an anti-cholesterol effect in a high-fat diet (HFD) in mice [20]. Analysis of the experimental results showed that the level of serum GT, GPT, and leptin, as well as the expression of hepatic COX-2 mRNA, as well as splenic COX-2 and IL-4 mRNA, were significantly higher in the HFD groups than in the control group (P > 0.05). With the exception of splenic IL-4 levels, these increases were significantly reduced by the addition of FCC. Expression of ICAM-1, a marker of aortic inflammation, was significantly increased in the HFD group. FCC suppressed weight gain and epididymal fat appendages, as well as inflammatory responses in the liver and spleen of HFD-fed mice.



Kazumi Sagayama, Naonobu Tanaka, Takatoshi Fukumoto, Yoshiki Kashiwada (2019) identified, after studying the effect of Inonotus obliquus on the prolif-eration of human papillary follicle cells (Hfdpc), five lanostane-type triterpenes (1-5) using spectroscopic data, of which lanosterol (1), inotodiol (3), lanost-8,24-diene-3β, 21-diol (4) and trametenolic acid (5) showed a more potent proproliferative effect on Hfpdc than minoxidil, an anti-alopecia agent used as a posi-tive control. Lanostane triterpenes (1, 3, 4, and 5) were potential candidates for new products that can be used for hair care with a stimulating effect on hair growth [27].

Devi K.S. and Maiti T.K. (2016), in a review of patents for the extraction of biologically active substances (BAS) from chaga mushroom, showed that for decades the use of components from mushrooms was considered in the framework of ethnic medicine and was widely used in the treatment of various serious diseases [6]. Methods of extraction of pharmacologi-cally significant fungal glucans affect their immunos-timulating properties, structure, composition, solubil-ity in water and conformation in solution. In addition, modifications of glucans depending on the particle size also contribute to a significant increase in their activity [15].

In order to increase the antitumor activity of polysaccharides and improve their clinical proper-ties, which depend mainly on their solubility in water, the main procedures used to optimize the isolation of polysaccharides are Smith decomposition (redox hydrolysis), formolysis, and carboxymethylation [31].

Gil Y.G., Kang S., Chae A. et al (2018) devel-oped and synthesized anisotropic porous palladium nanoparticles with full-wavelength absorption in the ultraviolet–visible–near-infrared range using concen-tration-dependent reduction synthesis to maximize the pharmacological activity of chaga mushroom (Inonotus obliquus) extract [9]. Porous Pd nanoparticles with chaga extract showed a surface antitumor effect, controlled delivery of doxorubicin via electrostatic interaction, and photothermal conversion effect under 808 Nm laser irradiation. The combined use of three approaches to cancer treatment has clearly demon-strated the feasibility of synergistic trimodal therapy. A modern platform using Pd, which is a key component of nanocatalysts, while not often used in biological applications, involves numerous applications using PD nanostructures, as well as the potential for develop-ment of new cancer treatments.

Kutaiba Ibrahin Alzand, Sabri Ünal, Mansor S. Mostafa Boufaris (2018) isolated three new lanostane-type triterpenes, a new abietane-type diterpenes, and 10 known compounds (5-14) from sclerotia of Inono-

tus obliquus [1]. Their structures were established using a combination of spectrometric methods, including infrared, 1-dimensional and 2-dimensional nuclear magnetic resonance and high-resolution electro-spray mass spectrometry. During in vitro studies, the compounds showed hepatoprotective effects against D-galactosamine-induced damage in WB-F344 cells with inhibitory effects ranging from 35.4% to 83.8%. Some of the compounds showed selective cytotoxicity against Bel-7402, A549 or KB cell lines and inhibitory action against protein tyrosine kinases with half-maximum inhibitory concentrations of 23.8 and 7.4 mmol/l, respectively.

A simpler way to prepare the extract is to extract it from crushed fruit bodies of the fungus by boiling them in water (0.5 l) for 30 minutes. The yield of the extract from mushrooms will be 30% on average.

In the same volume of ethanol, an alcohol extract is prepared from the crushed fruit bodies of the fungus I. obliquus (0.5 l) for 5 days. The yield of the extract from mushrooms is 60% on average. The absence of side effects of the use of I. obliquus ethanol extract (200 mg/kg) and of signs of toxicity or sensitivity in rats after oral administration of I. obliquus ethanol extract with anti-ulcer activity in all models used is due to the presence of various biologically active com-pounds [14].

C O N C L U S I O NWater and alcohol extracts of I. obliquus that

inhibit the proliferative activity of malignizing cells suggest that these fungi can potentially be used as an easily accessible source of natural antioxidants and in-hibitors of the process of carcinogenesis and metastasis with promising potential in the prevention of meta-bolic disorders associated with reactive oxygen species (ROS). I. obliquus should be considered as a source for the manufacture of drugs with antioxidant, antimicro-bial and antihyperglycemic activity. The most impor-tant component of chaga is betulinic acid, which ranks first in the ORAC for the level of antioxidants in food. The expressed activity of iron recovery dictates the study of the antiviral properties of the fungus I. obliq-uus, which is especially important at the present stage for the upcoming solution of issues with therapeutic and anti-epidemic measures against COVID-19. The results are encouraging, they explain and confirm the possibility of nutritional use of mushroom fruit bodies for the prevention and treatment of oxidative damage in neurodegenerative diseases, oncological pathology of tumors of various genesis and localization, as well as diseases of the pancreas and liver as hepatoprotec-tive agents, and also as anti-allergic agents with a mild sedative effect. Chaga mushroom extracts can prevent



the development of breast, ovarian, cervical, prostate, lung, stomach, spleen, brain, and thymus cancer in the initial stages, as well as leukemia, lymphoma, and melanoma.

The effectiveness of the drugs made from extracts of the birch chaga mushroom (Inonotus obliquus) on tumors of various localization is high and allows us to conceptually justify the clinical application for patients with cancer. Glucan and triterpenoid components of the fungus allow for the use of I. Obliquus in some cases as a direct antitumor agent and a regulator of apoptosis. The use of polysaccharides from I. obliquus can actually prevent tacrine-induced hepatotoxicity. Analysis of the results of numerous studies provides experimental evidence confirming the prospects for the use of I. obliquus in the treatment of lung cancer, and reveals the molecular basis underlying its cytotoxic activity against human cancer cells. Inotodiol ob-tained from I. obliquus can be used for the prevention and treatment of not only chronic conditions such as diabetes but also breast cancer.

R E F E R E N C E S1. Alzand K.I., Ünal S, Boufaris M.S.M. Lanos-

tane-Type Triterpenes and Abietane-Type Diterpene from the Sclerotia of Chaga Medicinal Mushroom, In-onotus obliquus (Agaricomycetes), and Their Biological Activities.//Int J Med Mushrooms. 2018;20(6):507–516. doi: 10.1615/IntJMedMushrooms.2018026007.

2. Arata S., Watanabe J., Maeda M., Yamamoto M., Matsuhashi H., Mochizuki M., Kagami N., Honda K., Inagaki M. Continuous intake of the Chaga mushroom (Inonotus obliquus) aqueous extract suppresses cancer progression and maintains body temperature in mice.//Heliyon. 2016 May 12;2(5):e00111. doi: 10.1016/j.heliyon.2016.e00111.

3. Baek J., Roh H.S., Baek K.H., Lee S., Lee S, Song S.S., Kim K.H. Bioactivity-based analysis and chemical characterization of cytotoxic constituents from Chaga mushroom (Inonotus obliquus) that induce apoptosis in human lung adenocarcinoma cells.//J Ethnopharmacol. 2018 Oct 5;224:63–75. doi: 10.1016/j.jep.2018.05.025.

4. Balandaykin M.E., Zmitrovich I.V.. Review on Chaga Medicinal Mushroom, Inonotus obliquus (High-er Basidiomycetes): Realm of Medicinal Applications and Approaches on Estimating its Resource Poten-tial.//Int J Med Mushrooms. 2015;17(2):95–104.

5. Chung MJ, Chung CK, Jeong Y, Ham SS. Anticancer activity of subfractions containing pure compounds of Chaga mushroom (Inonotus obliquus) extract in human cancer cells and in Balbc/c mice bearing Sarcoma-180 cells.//Nutr Res Pract. 2010 Jun;4(3):177–82. doi: 10.4162/nrp.2010.4.3.177.

6. Devi K.S. and Maiti.T.K. Immunomodulatory and Anti-cancer Properties of Pharmacologically Relevant Mushroom Glycans.//Recent Pat Biotechnol 2016 – Review. PMID 27456820

7. Dragendorff G. L. the Relationship between chemical components and Botanical features of plants" / / Pharmaceutical journal of Russia. – 1879.

8. Eid J.I., Das B. Molecular insights and cell cycle as-sessment upon exposure to Chaga (Inonotus obliquus) mushroom polysaccharides in zebrafish (Danio re-rio).//Sci Rep. 2020 May 4;10(1):7406. doi: 10.1038/s41598-020-64157-3.

9. Gil Y.G., Kang S., Chae A., Kim Y.K., Min D.H., Jang H. Synthesis of porous Pd nanoparti-cles by therapeutic chaga extract for highly efficient tri-modal cancer treatment.//Nanoscale. 2018 Nov 1;10(42):19810–19817. doi: 10.1039/c8nr07172a.

10. Hong K.B., Noh D.O., Park Y., Suh H.J. Hepato-protective Activity of Water Extracts from Chaga Medicinal Mushroom, Inonotus obliquus (Higher Basidiomycetes) Against Tert-Butyl Hydroperoxide-Induced Oxidative Liver Injury in Primary Cul-tured Rat Hepatocytes.//Int J Med Mushrooms. 2015;17(11):1069–76. DOI: 10.1615/intjmedmush-rooms.v17.i11.70

11. Hou Y.N, Deng G, Mao J.J. Practical Applica-tion of "About Herbs" Website: Herbs and Dietary Supplement Use in Oncology Settings.//Cancer J. 2019 Sep/Oct;25(5):357–366. doi: 10.1097/PPO.0000000000000403.

12. Isabel CFR, Lillian B, Rui MVA. Antioxidants in wild mushrooms.. Curr Med Chem. 2009; 16 (12):1543–1560.

13. Kang J.H., Jang J.E., Mishra S.K., Lee H.J., Nho C.W., Shin D., Jin M., Kim M.K., Choi C., Oh S.H. Ergosterol peroxide from Chaga mushroom (In-onotus obliquus) exhibits anti-cancer activity by down-regulation of the β-catenin pathway in colorectal cancer.//J Ethnopharmacol. 2015 Sep 15;173:303–12. doi: 10.1016/j.jep.2015.07.030.

14. Kikuchi Y, Seta K, Ogawa Y, Takayama T, Na-gata M, Taguchi T, Yahata K. Chaga mushroom-induced oxalate nephropathy.//Clin Nephrol. 2014 Jun;81(6):440–4. doi: 10.5414/CN107655.

15. Ko S.K., Jin M., Pyo M.Y. Inonotus obliquus extracts suppress antigen-specific IgE production through the modulation of Th1/Th2 cytokines in ovalbumin-sensitized mice.//J Ethnopharmacol. 2011 Oct 11;137(3):1077–82. doi: 10.1016/j.jep.2011.07.024.

16. Lee H.S., Kim E.J., Kim S.H. Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells.//Nutr Res Pract. 2015 Apr;9(2):111–6. doi: 10.4162/nrp.2015.9.2.111.

17. Lemieszek MK, Langner E, Kaczor J, Kandefer-Szerszeń M, Sanecka B, Mazurk-iewicz W, Rzeski W. Anticancer effects of fraction isolated from fruiting bodies of Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyl-lophoromycetideae): in vitro studies.//Int J Med Mushrooms. 2011;13(2):131–43.

18. Li Z., Mei J., Jiang L., Geng C., Li Q., Yao X., Cao J. Chaga Medicinal Mushroom, Inonotus



obliquus (Agaricomycetes) Polysaccharides Suppress Tacrine-induced Apoptosis by ROS-scavenging and Mitochondrial Pathway in HepG2 Cells.//Int J Med Mushrooms. 2019;21(6):583–593. doi: 10.1615/In-tJMedMushrooms.2019030857.

19. Mishra SK, Kang JH, Kim DK, Oh SH, Kim MK. Orally administered aqueous extract of Inonotus obliquus ameliorates acute inflammation in dextran sulfate sodium (DSS)-induced colitis in mice.//J Ethnopharmacol. 2012 Sep 28;143(2):524–32. doi: 10.1016/j.jep.2012.07.008.

20. Na H.G, Park Y., Kim M.A., Lee J.W., So G., Kim S.H., Jang K.H., Kim M.J, Namkoong S, Koo HJ, Lee SR, Sohn EH. Secondary Fermented Extract of Chaga-Cheonggukjang Attenuates the Effects of Obesity and Suppresses Inflammatory Response in the Liver and Spleen of High-Fat Diet-Induced Obese Mice.//J Microbiol Biotechnol. 2019 May 28;29(5):739–748. doi: 10.4014/jmb.1902.02034.

21. Nagajyothi PC, Sreekanth TV, Lee JI, Lee KD. Mycosynthesis: antibacterial, antioxidant and antipro-liferative activities of silver nanoparticles synthesized from Inonotus obliquus (Chaga mushroom) extract.//J Photochem Photobiol B. 2014 Jan 5;130:299–304. doi: 10.1016/j.jphotobiol.2013.11.022.

22. Nakajima Y, Nishida H, Matsugo S, Konishi T. Cancer cell cytotoxicity of extracts and small phenolic compounds from Chaga [Inonotus obliquus (persoon) Pilat].//J Med Food. 2009 Jun;12(3):501–7. doi: 10.1089/jmf.2008.1149.

23. Nguyen T.M.N., Lomunova M., Le B.V., Lee J.S., Park S.K., Kang J.S., Kim Y.H., Hwang I. The mast cell stabilizing activity of Chaga mushroom critical for its therapeutic effect on food allergy is derived from inotodiol.//Int Immunopharmacol. 2018 Jan;54:286–295. doi: 10.1016/j.intimp.2017.11.025.

24. Nguyen T.M.N., Le H.S., Le B.V., Kim Y.H., Hwang I. Anti-allergic effect of inotodiol, a lanostane triterpenoid from Chaga mushroom, via selective inhibition of mast cell function.//Int Immunophar-macol. 2020 Apr;81:106244. doi: 10.1016/j.int-imp.2020.106244.

25. Nikitina S.A., Khabibrakhmanova V.R., Sysoeva M.A. Composition and biological activ-ity of triterpenes and steroids from Inonotus obliquus (chaga).//Biomed Khim. 2016 May;62(4):369–75. doi: 10.18097/PBMC20166204369.

26. Ning X, Luo Q, Li C, Ding Z, Pang J, Zhao C. Inhibitory effects of a polysaccharide extract from the Chaga medicinal mushroom, Inonotus obliquus (higher Basidiomycetes), on the proliferation of hu-man neurogliocytoma cells.//Int J Med Mushrooms. 2014;16(1):29–36.

27. Sagayama K, Tanaka N, Fukumoto T, Kashi-wada Y. Lanostane-type triterpenes from the sclerotium of Inonotus obliquus (Chaga mushrooms) as proproliferative agents on human follicle dermal papilla cells.//J Nat Med. 2019 Jun;73(3):597–601. doi: 10.1007/s11418-019-01280-0.

28. Sun Y., Yin T., Chen X.H., Zhang G., Curtis R.B., Lu Z.H., Jiang J.H. In vitro antitumor activity and structure characterization of ethanol extracts from wild and cultivated Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromyc-etideae).//Int J Med Mushrooms. 2011;13(2):121–30.

29. Wang Q., Mu H., Zhang L., Dong D., Zhang W., Duan J. Characterization of two water-soluble lignin metabolites with antiproliferative activities from Inonotus obliquus.//Int J Biol Macromol. 2015 Mar;74:507–14. doi: 10.1016/j.ijbiomac.2014.12.044.

30. Wasser S.P. Current Findings, Future Trends, and Unsolved Problems in Studies of Medicinal.// Appl Microbiol Biotechnol Mar 2011. 89 (5), 1323–32.

31. Wasser S.P. Medicinal Mushroom Science: Current Perspectives, Advances, Evidences, and Challenges.//Biomed J Nov-Dec 2014. – 37 (6), 345–56.

32. Xin X., Qu J., Veeraraghavan V.P., Mohan S.K., Gu K. Assessment of the Gastroprotective Effect of the Chaga Medicinal Mushroom, Inonotus obliquus (Agaricomycetes), Against the Gastric Mu-cosal Ulceration Induced by Ethanol in Experimental Rats.//Int J Med Mushrooms. 2019;21(8):805–816. doi: 10.1615/IntJMedMushrooms.2019031154.

33. Yen-Nien Hou, Gary Deng, Jun J. Mao. From the Integrative Medicine Service, Memorial Sloan Ket-tering Cancer Center, New York, NY.// Cancer J 2019 Sep/Oct;25(5):357–366. Authors: Jun J Mao. Cancer J 2019 Sep/Oct;25(5):305–306. From the Memorial Sloan Kettering Cancer Center Bendheim Integrative Medicine Center, New York, NY.

34. Zhang P., Cao X., Li C., Zheng Z., Yong S., Jiang J.H. Cloning and Characterization of a Squalene Synthase Gene from the Chaga Medicinal Mushroom, Inonotus obliquus (Agaricomycetes).//Int J Med Mushrooms. 2016;18(5):445–55.

35. Zhang S.D., Yu L., Wang P., Kou P., Li J., Wang L.T., Wang W., Yao L.P., Zhao X.H., Fu Y.J. Inoto-diol inhibits cells migration and invasion and induces apoptosis via p53-dependent pathway in HeLa cells.//Phytomedicine. 2019 Jul;60:152957. doi: 10.1016/j.phymed.2019.152957.

36. Zhang X/., Bao C., Zhang J. Inotodiol suppresses proliferation of breast cancer in rat model of type 2 diabetes mellitus via downregulation of β-catenin sign-aling.//Biomed Pharmacother. 2018 Mar;99:142–150. doi: 10.1016/j.biopha.2017.12.084.



BIOCHEMICAL STUDY OF MENSTRUAL BLOOD AND A MATHEMATICAL MODEL FOR DIAGNOSIS OF UTERINE MYOMAS

A B S T R A C T — Uterine myomas or fibroids (UF) are the most common tumors in women, which can cause serious complications, often require surgical interference and cause infertility. The authors proposed a comprehensive approach to solving this issue, which includes identifying the characteristic structures of the solid phase of menstrual discharge and the introduction of a mathematical model for predicting the growth rates of UF. Patients were divided into three groups of women of reproductive age. The control group included 41 relatively healthy patients. The second group consisted of 40 women with simple UF (group 1), and 30 women with proliferating UF were included in the third group (group 2). Development and implementation of a method for diagnosing UF, which includes the study of morphological structures of solid phase of menstrual blood and levels of ligands inducing apoptosis and proliferation, allows to find the most optimal solution for this clinical task. A mathematical model is proposed for predicting the growth rates of UF. The safety and atraumatic method of morphological analysis of menstrual fluids makes it possible to apply it effectively in the work of an outpatient clinic.

K E Y W O R D S — uterine fibroids, proliferating uterine fibroids, proliferation, apoptosis, menstrual blood.

Yuliana Ukhanova , Adelya Zoeva ,Olga Tishkova , Svetlana Belyavskaya ,Adilya Ayupova , Pavel Loginov

Astrakhan State Medical University, Astrakhan, Russia

[email protected]

I N T R O D U C T I O NUterine myomas or fibroids (UF) are the most

common benign tumors of female genital organs in the world [1, 2, 3, 4], affecting at least 20–40% of women during their lifetime. Moreover, it is UF that accounts for one third to half of all hysterectomies. According to the results of a study carried out by a group of scientists headed by I.S. Sidorova in 2000, it was proposed to distinguish two clinical and morpho-logical types of the disease: simple and proliferating UF [3]. Moreover, at the same time, other researchers studied this aspect and came to the similar conclu-sions [1]. Simple and proliferating UF can vary on a number of features, such as, the location of nodes, their number, size, ultrasound characteristics.

Received 03 March 2021; Received in revised form 14 April 2021; Accepted 19 April 2021

http://dx.doi.org/10.35630/2199-885X/2021/11/2/G.1

At the cellular level, simple UF is characterized by low activity of proliferation and angiogenesis processes in combination with increased synthetic activity and severity of the stromal component [4]. At the same time, proliferating UF is characterized by rapid growth and high mitotic activity [5]. Biological fluids, such as menstrual blood, cervical mucus and endometrial lavage, contain markers that allow one to suspect and identify the presence of abnormalities in the organs of the reproductive system [1, 6, 7, 8, 10].

The development and implementation of a method for the diagnosis of UF, including the study of the morphological structures of the solid phase of menstrual blood and the levels of ligands inducing apoptosis (LIA) and proliferation (LIP), makes it pos-sible to find the most optimal solution to this clinical problem [9, 11].

The aim of the study was to develop a diagnostic method for proliferating uterine fibroids by determining the structural features of menstrual blood, identifying LIA and LIP in them, and to make a mathematical model for predicting the possible growth rate of UF.

M A T E R I A L A N D M E T H O D SWomen of reproductive age (23–45 years old)

were divided into three groups. The investigation was held at the Gynecology and Obstetrics Department of Astrakhan State Medical University. The control group included 41 relatively healthy women. The second group consisted of 40 women with simple UF (group 1), and the third group consisted of 30 women with proliferating UF (group 2). The investigation was held in accordance with the universal ethical stand-ards.

The distribution of women into groups with simple and proliferating UF, as well as the exclusion of women with «false» rapid growth was carried out on the basis of the results of histological examination of myoma nodes, ultrasound data, acute phase proteins, clinical picture and growth rates of myoma. The study included only patients with a «true» type of rapid UF growth. In the course of the work, data on the number, localization, and size of myoma nodes were analyzed, obtained by ultrasound performed on days 5–7 of the menstrual cycle.


https://orcid.org/0000-0001-8917-0089

https://orcid.org/0000-0001-8596-6514

https://orcid.org/0000-0002-1894-5809

https://orcid.org/0000-0003-2560-2449

https://orcid.org/0000-0001-9872-1401

https://orcid.org/0000-0001-5661-3648

http://dx.doi.org/10.35630/2199-885X/2021/11/2/G.1


Menstrual blood was taken in the most abun-dant days of menstruation. The obtained samples were centrifuged at 3000 rpm for 5 min. The resulting supernatants were used for further morphological and immunochemical studies. The supernatant was dropped onto a glass slide and covered with a coverslip. The material obtained was dried at room conditions for 2–3 days [12].

The analysis of the structures of the solid phase of menstrual fluid formed during the edge dehydra-tion was carried out using microscope Leica MZ 12.5 (Switzerland) equipped with a Leica ICC 50 digital camera.

The content of LIP and LIA in the menstrual blood supernatant was determined by enzyme immu-noassay using Bender MedSystems kits (Austria) [11].

The study results were processed using Microsoft Office 2010, Statistica software (StatSoft Inc., USA version 8.0). To study and compare the data obtained, the Kruskal-Wallis, Kolmogorov-Smirnov tests, as well as the method of correlation analysis were used. Description of qualitative features was reproduced in percent (%), absolute values (n/N) and contingency tables with the definition of χ2; description of quantita-tive data was realized in the form of M ± m [11]. The results were considered reliable at p < 0.5.

R E S U L T SIn group 1, the nodes were located mainly in the

fundus and body of the uterus, which was recorded in 26 patients and accounted for 65.0% of observations (p < 0.05). In group 2, the nodes were located intrali-gamentally (in 40.0% of patients), in the area of fundus and body of the uterus (in 43.3% of women), as well as in the isthmus (in 16.7% of patients). The number of myoma nodes with simple UF varied from 1 to 4 and averaged 1.6 ± 0.14, while with proliferating UF there was a significant increase in the number of nodes to 5–7, with average number of 4.0 ± 0.14 (p<0.05). The size of the dominant node with simple UF was 2.6±0.19 cm, with proliferating UF 5.7±0.48cm (p<0.05).

It has been proved that the presence of parallel structures in the morphological picture of the biologi-cal fluid indicates the presence of hyperplastic proc-esses in patients [12, 13]. In this regard, a significant increase in their percentage in group 2 confirmed the predominance of proliferation processes in this contingent [11]. At the microscope level, proliferation markers — parallel (Fig. 1) and fibrous (Fig. 2) struc-tures are seen in patients with endometrial hyperplastic processes.

The conducted immunochemical study showed that in menstrual blood of the control group, the

level of LIP was 3.0–4.8 ng/ml, in group 1 it rised to 4.8–6.4 ng/ml, and in group 2 up to 11.1–14.7 ng/ml. In turn, LIA in the control group was in the range of 35.1–36.8 pg/ml, in group 1 it decreased to 24.0–27.0 pg/ml, and in group 2 up to 20.0–22.5 pg/ml [11, 14, 15].

Correlation analysis in the group of patients with simple UF (group 1) revealed a relationship between lattice structures and levels of LIP (r = +0.3) and LIA (r = +0.28), transitional forms and concentration of LIP (r = +0.27) and dendritic forms and LIA (r = +0.3) [11]. In patients with proliferating MM (group 2), there was a direct relationship between the LIP level and the size of uterus (r = +0.35) according to the ultrasound examination of pelvic organs [15]. The results of the investigation are summarized in Table 1.

At the final stage, to derive a mathematical for-mula, the binary logistic regression method was used:

,

where: P is the probability of the event occurring; e — constant value (2.72), which serves as the base of natural logarithm; z — the degree of inverse logarithm is calculated by the formula:

z = 2.172 FS + 2.238 PS + 1.568 NN – 10.915

where: FS — fibrous structures, PS — parallel struc-tures, NN — the number of nodes [16].

In the case when P was more than 0.5, the risk of developing proliferating UF increased (more than 50%). In this group of patients, at the next stage, the levels of LIA and LIP were analyzed. With an increase in LIP of more than 11.1 ng/ml and a decrease in LIA of less than 22.5 pg/ml, the risk of an event tends to approach 100% [11].

The introduction of this model into clinical practice will enable, with a high degree of probability, to make a prognosis of the proliferative potential of myoma nodes for each individual patient, which, in turn, gives an answer to the question of the advisability of using hormone therapy in the pre- and postopera-tive period in women who need myomectomy.

Patients with a high risk of developing proliferat-ing UF, for whom myomectomy is indicated, need either isolated or combined pre- and postoperative treatment with the drug class of gonadotropin-releas-ing hormone agonists in order to suppress proliferative processes. At the same time, for patients with simple UF, a one-stage treatment, including myomectomy, is sufficient, followed by a decision on planning preg-nancy.



C O N C L U S I O NTherefore, identification of characteristic struc-

tural features of menstrual blood in patients with different clinical and morphological variants of the tu-mor, along with the determination of levels of LIP and LIA, allows not only to make an individual prognosis, to plan the management and treatment of patients, but also ultimately helps achieve the main goal — the im-plementation of reproductive life planning (RLP). At the same time safety and simplicity of morphological method of biological fluids makes it possible to apply actively this method in an outpatient care facility.

R E F E R E N C E S1. Ciavattini A., Clemente N., Delli C.G.,

Di G.J., Giannubilo S.R., Tranquilli A.L. (2015). Number and size of uterine fibroids and obstetric outcomes. The Journal of Maternal-Fetal & Neonatal Medicine, 28(4): 484–488. DOI: 10.3109/14767058.2014.921675

2. Dikareva L.V., Shvarev Е.G., Biryukova A.F. (2012). Biochemical features of the endometrial secre-tion of patients with uterine fibroids. Astrakhanskiy meditsinskiy zhurnal, 7(2), 175–178. (in Russ.)

3. Sidorova I.S., Hunanyan A.L., Kogan E.A., Guriev T.D. (2010). Uterine fibroids in young pa-tients: clinical and pathogenetic features. Akusherstvo, ginekologiya, reprodukciya, 4(1), 16–20.

4. Buyanova S.N., Yudina N.V., Gukasyan S.A., Mgeliashvili M.V. (2012). Modern aspects of the growth of uterine fibroids. Rossiiskii vestnik akushera-ginekologa, 4, 42−48. (in Russ.)

5. Dikareva L.V., Shvarev E.G., Ayupova A. K. (2008). Clinical and diagnostic aspects of fast-growing uterine fibroids. Problemy reproduktsii, 14(6):20–25.

6. Dikareva L.V., Shvarev E.G., Ovodenko D.L., Shramkova I.A., Arutyunyan A.A. (2008). Method for diagnosing endometrial pathology in patients with uterine fibroids. Patent RF, no. 2334987.

7. Buyanova S.N., Logutova L.S., Babunashvili E.L., Gorbunova T.N. (2003). Reproductive prog-nosis in uterine fibroids. Rossiiskii vestnik akushera-ginekologa, 4: 47−49. (in Russ.)

8. Dikareva L.V., Shvarev E.G., Ayupova A.K. (2008). Clinical and prognostic significance of the rap-idly growing uterine fibroids. Uspehi sovremennogo estestvoznanija, 8: 102–105. (in Russ.)

Fig. 1. Parallel structures in an analytical cell. Magnification 100× Fig. 2. Fibrous structures in an analytical cell. Magnification 100×

Isotropic forms Control group (n = 41), % Group 1 (n = 40), % Group 2 (n = 30), %Dendritic 41,5 47,5 13,31Transitional 34,2 45,0 3,31Lamellar 9,8 27,52 3,31Parallel 2,4 37,52 83,31Fibrous 0 10,0 50,01

Table 1. Structural elements of the solid phase of the menstrual blood

Note: 1) p < 0.05 — significant differences compared to group 1; 2) p < 0.05 — significant differences compared with the control group



9. Dikareva L.V., Ajupova A.K., Shvarev E.G., Ukhanova Ju.Yu., Zoeva A.R., Gadzhieva P.H. Diagnostic aspects of obstetric and gynecological pathology of solid phase morphology of biological liq-uids (review). Estestvennye nauki, 3: 40–48. (in Russ.)

10. Dikareva L.V., Shvarev E.G., Shramkova I.A., Shvarev G.E. (2008). A method for diagnosing a rapid-growing uterine myoma. Patent RF, no. 2332167.

11. Ukhanova Ju.Yu. Markers of proliferation and apop-tosis in the diagnosis of proliferating uterine fibroids. (2017). Aktualnyye voprosy sovremennoy medisiny, 192−194. (in Russ.)

12. Shabalin V.N., Shatohina S.N. (2001). Тhe mor-phology of the human biological fluids (303−303). Hrizostom Publ. (in Russ.)

13. Shatohina S.N., Shabalin V.N. (2004). Мethod of diagnosing a cell hyperplasia organism. Patent RF, no. 2234856.

14. Stewart E.A., Cookson C.L., Gandolfo R.A., Schulze-Rath R. (2017). Epidemiology of uterine fibroids: a systematic review. BJOG: An International Journal of Obstetrics & Gynaecology, 124(10):1501–1512. DOI: 10.1111/1471-0528.14640

15. Ukhanova Ju.Yu., Dikareva L.V., Shvarev E.G., Ajupova A.K. (2015). An integrated approach to the diagnosis of uterine fibroids and assessment of its growth rate. Tumors of female reproductive system, 11(4):71–75. (in Russ.)

16. Ukhanova Ju.Yu., Dikareva L.V., Shvarev E.G., Ajupova A.K. (2015). An innovative approach to the diagnosis of fast-growing uterine fibroids. Astra-khanskiy meditsinskiy zhurnal, 10(3):106−114. (in Russ.)



THE RESULTS OF USING A BIOACTIVE GLASS-BASED COATING BY DEPOSITION ON THE CONTACT SURFACE OF PLATES IN BONE FRACTURES ASSOCIATED WITH TUMORS

A B S T R A C T — The article reports on the results of surgical treatment in 20 patients with bone tumors using a coating containing bioactive glass on the contact surface of the fixation plate. As a result of metallic osteosynthesis of the bones, infectious complications were observed in 1 (5%) patient and tumor recurrence was observed in 2 (10%) patients. Limb function after metallic osteosynthesis ranged from 72.3% to 97.4% depending on the location of the lesion focus. The quality of life of the operated patients increased to 75–85 points. Thus, the use of plates with bioactive glass-based material deposited on the contact surface of the plate, in metallic osteosynthesis for pathological bone fracture or at the risk of pathological bone fracture achieves more effective integration of metal plate with bone. Consequently, a better functional stability of the bone is achieved at the fracture site. Morphological examination of bone biopsy samples from the implantation site of the plate and the plate itself with the deposited bioactive glass-based coating revealed active regeneration of bone tissue. This resulted in an increase in the density of the restored bone. According to the results of histological and morphometric examination it can be stated that deposition of a material containing bioactive glass on the contact surface of the metal plate promotes reparative osteogenesis at the site of bone damage and its morphogenesis of the lamellar type.

K E Y W O R D S — metallic osteosynthesis, plate, bioactive glass-based (BG) coating, bone tumor, morphological study.

Oleksandr Burianov¹ , Volodymyr Protsenko² ,Audai Abudayeh ¹, Volodymyr Chornyi¹, Volodymyr Konovalenko³, Yevhen Solonitsyn²

1 Bogomolets National Medical University, Kyiv2 Institute of Traumatology and Orthopedics, National Academy of Sciences of Ukraine, Kyiv3 R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, Kyiv, Ukraine

[email protected]

ments, optimal rate and rhythm of distraction, sparing attitude to osteogenic tissue, good blood supply to the operated limb, the possibility of functional use of the limb, starting from the first days after surgery, represent conditions for reparative regeneration of bone tissue [1, 2]. A study of the reference literature shows the application of autogenous grafts of bone tissue and various bioactive products of decalcified bone and biocomposite matrices, recombinant human bone morphogenetic proteins and ceramics. All the materials including new cellular technologies are used to stimulate osteogenesis, [3, 4, 5]. Over recent years, there has been an active trend in the development of biomaterials aimed at creating composites that replace damaged tissues including bone tissue [6]. In orthoped-ics, this was facilitated by the development of the major joint arthroplasty industry and the use of bioceramic implants. [7, 8, 9]. Biomaterials used as implants or as temporary fixators for broken bone (periosteal plates, intramedullary nails) can be represented by biotolerant materials (stainless steel and cobalt-chromium alloys), bioinert (titanium and aluminum oxides), bioactive (calcium phosphate ceramics and silicon-based bioglass) [7]. There are no bioactive metals that would accelerate reparative osteogenesis. In orthopedics, chromium-nickel and chromium-nickel-molybdenum corrosion-resistant steels, alloys of cobalt, tantalum, titanium, pure metals — nickel, silver, titanium are most often used for the manufacture of surgical implants [10].

Typical representatives of bioactive materials in-clude bioglass (the most commonly used composition is 24.5% Na2O, 24.5% CaO, 45.0% SiO2, 6% P2O5, var-ying the composition, you can change their bioactivity and resorption) and materials based on hydroxyapatite (HA) — Ca10 (PO4)6(OH)2 (dense and porous ceram-ics) [11, 12]. Hydroxyapatite (Ca10(PO4)6(OH)2) is one of the few bioactive materials that supports bone ingrowth and osseointegration when used in ortho-pedic implants due to its high biocompatibility.

In the last few years, a special term has appeared in the literature – biocompatible nanoceramics, which means nanostructured ceramic material used in medicine to restore (replace) damaged hard tissues [9, 13]. The creation of composite materials based on

Received 30 January 2021 Received in revised form 04 March 2021; Accepted 15 March 2021

I N T R O D U C T I O NIn the treatment of fractures and diseases of the

skeletal system, internal (intramedullary and perio-steal) and external (pins and nails) fixators are used to ensure the most reliable osteosynthesis of bone. Complete reposition and stable fixation of bone frag-

S U R G E R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/11

https://orcid.org/0000-0002-2174-1882

https://orcid.org/0000-0001-8896-8371

https://orcid.org/0000-0001-7057-1405

http://dx.doi.org/10.35630/2199-885X/2021/11/2/11


biphasic ceramics using various binding components, biologically active substances that would provide oste-oinduction of osteoplastic material to form a matrix, on which bone tissue will be formed, is one of the promising areas [14]. The concept of biphasic com-posite materials in the system “HA – TCF (tricalcium phosphate)” was developed based on the assumption of the possibility of regulating the kinetics of biodeg-radation by changing the ratio: less (HA) and more (TCF) of soluble phases in one material. Dissolution of the TCF component in body fluid promotes the process of mineralization, and the biological behavior of biphasic ceramics depends on the ratio of HA/TCF [15]. The bioactivity of materials is determined mainly by chemical factors, such as the crystalline phase and molecular structure of the material, as well as physical factors — roughness and porosity of the material sur-face and the ability to form a chemical bond with the surrounding bone [8, 11]. A number of complex and closely interrelated processes take place on the surface of a bioactive implant. The surface of the material, its biocompatibility is closely related to the adhesion of osteogenic and mesenchymal stem cells on its surface [16, 17]. It is the adhesion as well as the distribution of these cells that will affect their ability to prolifer-ate and differentiate into osteoblasts upon contact with the implant. The latter is crucial in the process of establishing a mechanically strong interface with complete fusion between the implant surface and bone tissue without a layer of fibrous tissue, which is usually formed on the surface of a bioinert metal implant [18, 19, 20, 21]. In this article we report on the use of titanium plates in bone fractures on the background of tumors with a BG-based material, deposited on their contact surface.

Objective:To show the possibilities of application of

periosteal plates with a bioactive glass containging material deposited on the contact surface to improve integration of the plate with bone in pathological bone fractures associated with tumors.

M A T E R I A L A N D M E T H O D SDuring the period from 2017 to 2020, in Insti-

tute of Traumatology and Orthopedics of NAMS of Ukraine 20 operations on metallic osteosynthesis were performed using periosteal plates with a bioac-tive glass-based coating deposited on the contact surface of the plate, in pathological fractures or at risk of pathological fractures secondary to tumors of long tubular bones. The average age of patients was 39.2±1.4 years (from 28 to 57 years), among them there were 13 (65%) women and 7 (35%) men.

Metallic osteosynthesis was performed using periosteal plates of different lengths and configura-tions. Plates (medical steel grade BT6) had the fol-lowing composition: titanium as the main substance, aluminum content — 3.9/2.8 nm, manganese content — 1.3/1.2 nm, iron content — 0.4/0.1 nm, silicon content — 0.1/0.1 nm, calcium content on the surface of the plate — 0.3 nm, chlorine content — 0.2 nm, sulfur content — 0.1 nm.

Table 1 presents the morphological types of tumors identified during metallic osteosynthesis us-ing plates with a BG-based coating deposited on the contact surface.

Table 1. Morphological types of tumors identified during metallic osteosynthesis using plates with a BG-based coating deposited on the contact surface

Morphological types of tumors Number of cases %Metastatic tumors 14 70Chondroma 3 15Giant cell tumor of bone 1 5Solitary myeloma 1 5Lymphosarcoma of bone 1 5Total 20 100

The scope of surgery depended on the location of the tumor: patients underwent intraosseous bone resection with the tumor; bone defect in chondroma and giant cell tumor was filled with allograft or mate-rial containing bioactive glass; in metastatic tumor, solitary myeloma and lymphosarcoma bone defect was filled with polymethyl methacrylate, after which me-tallic osteosynthesis was produced using a BG-based coating deposited on the contact surface.

Table 2 presents location and number of cases of metallic osteosynthesis of bones using a BG-based ma-terial deposited on the contact surface of the implant.

The use of plates with a bioactive glass-based coating deposited on the contact surface achieves a better integration of the plates with the bone, as well as the fact that in bone tumors, the plates are usually not removed over time.

The material deposited on the contact surface of the plate was an implant material containing bioactive glass — Syntekist Biocomposite (certificate of state reg-istration No. 3653/2005 dated January 28, 2005), which was synthetically produced in the laboratory of the Institute of Materials Science of the National Academy of Sciences of Ukraine by the team of prof. V.A. Dubok.

Implant material — Syntekist biocomposite is a multiphase inorganic material synthetically produced

S U R G E R Y


by chemical precipitation and ceramic technology. Its phase composition includes bioactive glass — 50–65 wt.%, hydroxyapatite — 14–17 wt.%, whit-lockite — 14–17 wt.%, wollastonite — 7–9 wt.%. The chemical composition is shown in Table 3 (in terms of oxides).

The functional result of the operated limb was determined according to the MSTS Rating scale (Musculo-Sceletal Tumor Staging /System/). Patients' quality of life was determined in points according to the EORTC QLQ-30 questionnaire. Patient survival was determined using the Kaplan-Meier method.

F I N D I N G SOf 20 patients, infectious complications were

observed in 1 (5%) patient and tumor recurrence was observed in 2 (10%) patients after metallic osteo-synthesis for pathological fractures and the risk of pathological bone fractures, In a patient with an infec-tious complication, after removal of the plate, radical surgical treatment of the wound was performed with removal of all pathologically changed tissues, inocula-tion of fluid for culture from the place of installation of a plate, washing the wound with antiseptic solutions and installing an active drainage system with antibiotic irrigation.

In 2 patients with tumor recurrence a bone biopsy was performed after plate removal at the point of con-tact with the BG-based coating deposited on the plate. A biopsy of the metal plate surface was performed followed by morphological microscopic examination

Table 2. Location and number of cases of metallic osteosynthesis of bones using a BG-based material deposited on the contact surface of the implant

Location Number of cases %Distal femur 7 35Humeral diaphysis 4 20Tibial shaft 3 15Proximal tibia 2 10Distal tibia 1 5Proximal femur 1 5Radial shaft 1 5Femoral shaft 1 5Total 20 100

Table 3. Chemical composition of Syntekist biocomposite

CompositionNa2O K2O SiO2 CaO P2O5 Ag2O15,54 0,173 27,58 35,45 21,27 -

Range of permissible composition 15–17 0,1–0,3 25–28 34–37 20–23 0,01–0,08

It is a bioactive and osteoconductive biomaterial, which is available in the form of powders, granules, blocks and implants of complex shape with different activity, dispersion and adsorption capacity, with a sig-nificant range of porosity and mechanical properties. The contact surface of the plate was covered with a BG-based material by means of gas detonation deposi-tion. Abrasive treatment of the contact surface of the plate was performed in the gas detonation deposition unit before depositing the BG-based coating; on the surface a surface relief of a given depth was formed, which was equal to the size of osteon and provided a large contact area and adhesion strength between the bone and the plate. Next, the deposition of a layer of the BG-based material has been carried out, which allows to solve the problem of amorphization of the coating. Due to the presence of bioactive compounds in the coating, evenly distributed throughout its thickness, in the process of resorption of the coating in the body, it caused antibacterial, osteoconductive, antiresorptive and other effects throughout the life of the plate (see Fig. 1).

of the obtained material. These 2 patients subsequently underwent resection of bone articular segment and joint endoprosthesis replacement due to tumor recur-rence.

Functional outcome in the limb (according to the MSTS scale) after metallic osteosynthesis of the distal femur was 82.5%; in the humeral diaphysis — 95.4%, of the tibial shaft — 91.6%; in the distal tibia amount-ed to 86.8%; in the proximal tibia — 93.2%, in the proximal femur — 72.3%; in the radial shaft — 97.4%; in the femoral shaft — 76.4%.

The quality of life of patients according to the EORTC QLQ-30 questionnaire before metallic osteosynthesis was estimated as 20–30 points, after metallic osteosynthesis it was 75–85 points.

The overall three-year survival of the patients was 46.5 ± 2.9%, and the five-year survival was 26.2 ± 4.1%.

Here is an example from our practice: Patient Ya., 40 years old, considers herself ill since October 2020, when pain and swelling appeared in the distal segment of the left lower leg. According to the patient, the pain intensified within a month, so the patient was forced

S U R G E R Y


to walk on crutches. A month later, the patient went to the clinic of the State Institution "ITONAMNU", where X-ray examination revealed a tumor of the left distal tibia (see Fig. 2). In November 2020, the patient underwent an open biopsy of the lesion focus in the distal part of the left tibia. Histopathological find-ings: giant cell tumor of bone. In December 2020, the patient underwent a intraosseous resection of the lesion focus in the distal part of the left tibia with a tumor and a plastic repair of the defected femur using a BG-based material and extramedullary metallic osteosynthesis of the distal tibia using plate with a BG-based coating by deposition on its contact surface (see Fig. 3). No complications were observed in the postoperative period. The function of the left lower leg (according to the MSTS scale) was 86.8%. There was no tumor recurrence on X-ray in 1.5 months.

In 2 patients with tumor recurrence after plate removal (in 2 months after implantation) during the morphological examination of bone tissue from the area of plate removal and the surface of the metal plate coated with BG-based material we found out: in the subject No.1 bone perimeter was characterized by active osteogenesis. Crystalline granules were detected, which probably represent biocomposite residues after plate removal. Almost the entire surface of the plate was surrounded by newly formed bone tissue, mainly vowen bone. Although in some areas the morphogen-esis of lamellar bone tissue has been observed in newly formed trabeculae (Fig. 4). The relative surround-ing of the bone defect (at the site of bone trepana-tion) by newly formed bone tissue amounted to 82.0 (78.3–91.7)%.

In the subject No.2 during morphological ex-amination at the site of the defect, a damaged hyaline cartilage of the articular surface and the metaphyseal plate with replacement by connective tissue was re-vealed. In the areas where the granules of the biocom-posite got into the lacunae of the cortical bone, active

Fig. 1. A metal plate with a BG-based coating deposited on the contact surface

Fig. 2. Photographs of radiographs (A is a frontal and B is a lateral projection) of the left tibia of patient Ya. with a giant cell tumor of the distal tibia

Fig. 3. Photographs of radiographs (A is a frontal and B is a lateral projec-tion) of the left tibia of the patient Ya. after metallic osteosynthesis for a giant cell tumor of the distal tibia using plate with a BG-based coating deposited on its contact surface. The defect of the bone was repared and filled with the BG containing material

osteogenesis was detected around this material (Fig. 5). The biocomposite was partially surrounded by con-nective tissue, mostly an osseous one. Morphogenesis corresponded to the formation of lamellar bone tissue. The relative density of the newly formed bone tissue around the granules of the biocomposite reached 89.6 (72.6–97.8)%.

In the subject No.3 an epiphyseal defect was revealed during the morphological examination, and

S U R G E R Y


Fig. 4. Active osteogenesis around the metal plate. Note: d — defect; bt — bone tissue. Hematoxylin-eosin, vol. 20, approx. 10

Fig. 5. Active osteogenesis around the biocomposite. Note: b — biocom-posite; ct — connective tissue; bt — bone tissue. Hematoxylin-eosin, vol. 10, approx. 10.

d

bt

ct

bt

b

the centers of active osteogenesis around the granules of the biocomposite were detected in the cavities of the trabecular bone (Fig. 6). Almost the entire surface of the granules (92.5 (83.5–95.6)%) was surrounded by newly formed bone tissue (the lamellar type domi-nated). Isolation of the biocomposite from the sur-rounding morphofunctional bone formations (bone marrow, cartilaginous tissue of the articular surface, metaphyseal plate) was observed.

In the subject No.4 during the morphological examination it was detected that the bone defect was filled with connective tissue. Foci of inflammatory infiltration (neutrophils and macrophages) were de-

tected. The perimeter of the defect was surrounded by newly formed bone tissue, but the boundary between the defect and the bone tissue remained partially filled with connective tissue (Fig. 7).

Fig. 6. Active osteogenesis around the biocomposite. Note: b — biocom-posite; bt — bone tissue. Hematoxylin-eosin, vol. 10, approx. 10.

b

bt

Fig. 7. The bone defect is filled with connective tissue with macrophage infiltration. Note: defect; bt — bone tissue; ct — connective tissue. Hematoxylin-eosin, a: vol. 4, approx. 10

ct

bt

ct

In the subject No.5, a morphological examina-tion revealed an area of bone defect with biocomposite granules. Osteogenesis around the biocomposite gran-ules was uneven: one part was surrounded by connec-tive tissue, and the other part was totally integrated into regenerated bone tissue (Fig. 8). Newly formed bone tissue in most areas is lamellar.

During morphological examination in the subject No.6 (Fig. 9) the defect was detected at the level of epiphyse, filled with connective tissue, articular and

S U R G E R Y


metaphyseal hyaline cartilage are deformed. The metaphyseal plate is penetrated by a defect surrounded by newly formed bone tissue, which is lamellar to a greater extent. The level of regeneration was 87.2 (86.7–88.1)%.

R E S U S T SAs a result of metallic osteosynthesis in patho-

logical bone fracture or at risk of pathological bone fracture associated with bone tumors, positive treat-ment results were obtained: limb function ranged from 72.3% to 97.4% depending on the location of the lesion focus, quality of life of operated patients in-creased to 75–85 points. Morphological examination of bone biopsy material from the site of implantation

of the plate and from the plate itself with the deposited BG-based material revealed active regeneration of bone tissue leading to an increase in the density of the restored bone. The results of histological and morpho-metric examination demonstrate that the deposition of the BG-based material onto the contact surface of the metal plate promotes reparative osteogenesis at the site of bone damage and its morphogenesis of the lamellar type.

C O N C L C U S I O N1. As a result of application of periosteal plates

with deposition of the BG-based material for patho-logical bone fractures we observed infectious compli-cations in 1 (5%) patient and tumor recurrence in 2 (10%) patients, out of 20 patients with bone tumors.

2. The results of histological and morphometric examination showed that the deposition of the BG-based material onto the contact surface of the metal plate for metallic osteosynthesis promotes more effec-tive integration of the plate with bone, which leads to a greater functional stability of the bone at the fracture site.

R E F E R E N C E S1. Perren SM, Fernandez Dell'oca A, Regazzoni

P. Evolution of AO Fracture Treatment Part 1: the Internal Fixator. Acta Chir Orthop Traumatol Cech. 2017;84(6):413–417. English. PMID: 29351522.

2. Shevtsov VI, Shved SI, Sysenko YuM. Transos-seous fixation for treatment of fragmented fractures. Kurgan, 2002. 331 p. (in Russian).

3. Barinov S.M. Calcium phosphate-based ceramic and composite materials for medicine. Russian Chemical Reviews (2010),79(1):15–32 (in Russ.). Doi: 10.1070/RC2010v079n01ABEH004098.

4. Mironov S.P., Gintsburg A.L., Es'kin N.A., Lunin V.G., Kariagina A.S., Zaitsev V.V. Oste-oinductive implants based on biocomposite matrices and recombinant bone morphogenetic proteins (rh-BMP). Status, the prospect of using in traumatology and orthopaedics /Collected abstracts of IX Congress of traumatologists-orthopedists. Saratov, 2010;Vol.3: P. 1122–1123. (in Russian)

5. Shvedovchenko I.V., Shestakov V.P., Lebedeva N.N., Nikitchenko I.I., Svintsov A.A. Disability due to injuries and diseases of the locomotor system, and the ways of its prevention in the Russian Federa-tion. Collected abstracts of IX Congress of traumatol-ogists-orthopedists. Saratov, 2010. S. 1043–1044. (in Russian).

6. Auclair-Daigle C, Bureau MN, Legoux JG, Yahia L. Bioactive hydroxyapatite coatings on polymer composites for orthopedic implants. J Biomed Mater Res A. 2005 Jun 15;73(4):398–408. doi: 10.1002/jbm.a.30284. PMID: 15892136.

Fig. 8. Heterogeneous osteogenesis around the biocomposite. Note: b — biocomposite; ct — connective tissue; bt — bone tissue. Hematoxylin-eosin, vol. 20, approx. 10

Fig. 9. The defect is filled with connective tissue, active osteogenesis is present. Note: defect; bt — bone tissue; ct — connective tissue. Hematoxylin-eosin, vol. 4, approx. 10

b

bb

b

ct bt

ct

bt

bt

S U R G E R Y


7. Popkov A.V., Popkov D.A. Bioactive implants in traumatology and orthopaedics. Irkutsk: NTsRVKh SO RAMN, 2012. 434 p. (Russian).

8. Okada Y, Kobayashi M, Neo M, Kokubo T, Nakamura T. Ultrastructure of the interface be-tween bioactive composite and bone: comparison of apatite and wollastonite containing glass-ceramic filler with hydroxyapatite and beta-tricalcium phosphate fillers. J Biomed Mater Res. 2001 Oct;57(1):101–7. doi: 10.1002/1097-4636(200110)57:1<101::aid-jbm1147>3.0.co;2-g. PMID: 11416855.

9. Makkar P, Sarkar SK, Padalhin AR, Moon BG, Lee YS, Lee BT. In vitro and in vivo assess-ment of biomedical Mg-Ca alloys for bone implant applications. J Appl Biomater Funct Mater. 2018 Jul;16(3):126–136. doi: 10.1177/2280800017750359. Epub 2018 Apr 2. PMID: 29607729.

10. Stephenson PK, Freeman MA, Revell PA, Ger-main J, Tuke M, Pirie CJ. The effect of hydroxya-patite coating on ingrowth of bone into cavities in an implant. J Arthroplasty. 1991 Mar;6(1):51–8. doi: 10.1016/s0883-5403(06)80157-9. PMID: 1849985.

11. Eralp L, Kocaoğlu M, Ozkan K, Türker M. A comparison of two osteotomy techniques for tibial lengthening. Arch Orthop Trauma Surg. 2004 Jun;124(5):298–300. doi: 10.1007/s00402-004-0646-9. Epub 2004 Mar 18. PMID: 15034725.

12. Jones JR. Review of bioactive glass: from Hench to hybrids. Acta Biomater. 2013 Jan;9(1):4457–86. doi: 10.1016/j.actbio.2012.08.023. Epub 2012 Aug 21. PMID: 22922331.

13. Bauer TW, Muschler GF. Bone graft materials. An overview of the basic science. Clin Orthop Relat Res. 2000 Feb;(371):10–27. PMID: 10693546.

14. Lalan S, Pomerantseva I, Vacanti JP. Tissue engineering and its potential impact on surgery. World J Surg. 2001 Nov;25(11):1458–66. doi: 10.1007/s00268-001-0131-3. PMID: 11760750.

15. Olszewska-Słonina DM, Drewa TA, Styczyński J, Czajkowski R. Hodowla komórek, inzynieria tkankowa i medycyna regeneracyjna. Cześć II [Tissue engineering yesterday and today. Part II]. Wiad Lek. 2006;59(9-10):732-7. Polish. PMID: 17338141.

16. Anselme K, Bigerelle M. On the relation between surface roughness of metallic substrates and adhesion of human primary bone cells. Scanning. 2014 Jan-Feb;36(1):11–20. doi: 10.1002/sca.21067. Epub 2012 Nov 30. PMID: 23203601.

17. Ripamonti U. Functionalized Surface Geometries Induce: "Bone: Formation by Autoinduction". Front Physiol. 2018 Feb 6;8:1084. doi: 10.3389/fphys.2017.01084. PMID: 29467661; PMCID: PMC5808255.

18. Hench LL. The future of bioactive ceramics. J Mater Sci Mater Med. 2015 Feb;26(2):86. doi: 10.1007/s10856-015-5425-3. Epub 2015 Feb 3. PMID: 25644100.

19. Song HR, Myrboh V, Oh CW, Lee ST, Lee SH. Tibial lengthening and concomitant foot deformity correction in 14 patients with permanent deformity after poliomyelitis. Acta Orthop. 2005 Apr;76(2):261–9. doi: 10.1080/00016470510030670. PMID: 16097554.

20. Holzapfel BM, Rudert M, Hutmacher DW. Gerüstträgerbasiertes Knochen-Tissue-Engineering [Scaffold-based Bone Tissue Engineering]. Or-thopade. 2017 Aug;46(8):701–710. German. doi: 10.1007/s00132-017-3444-0. PMID: 28725934.

21. Paley D. Surgical reconstruction for fibular hem-imelia. J Child Orthop. 2016 Dec;10(6):557–583. doi: 10.1007/s11832-016-0790-0. Epub 2016 Dec 1. PMID: 27909861; PMCID: PMC5145840.

S U R G E R Y


RESULTS OF SURGICAL TREATMENT OF PATIENTS WITH PANCREATIC INJURIES

A B S T R A C T — The analysis of 12 cases of surgical treatment of young male patients, with injuries of the pancreas is presented. By the nature of the injuries, half of the patients had gunshot injuries, and 4 had blunt injuries, 2 patients with stab and slash wound. All injuries were combined; with severity grades I–III according to AAST classification. All patients underwent laparoscopic procedures. In one patient distal pancreatectomy was performed, whereas in 7 patients the appropriate sutured closure and in 4 patients drainage closure were performed. In one patient, we used a gastric transplant on a vascular pedicle. This case is of particular interest, so we presented it separately.

K E Y W O R D S — pancreas, surgery, injury, laparoscopy.

Givi Odishelashvili , Sergei Shashin ,Dmitri Pakhnov


[email protected]

I N T R O D U C T I O NAccording to various authors, the lesion on

the pancreas is quite rare and occupies the last place along with intestines, spleen, liver and kidneys injuries which represents less than 5% of abdominal trauma [1,2,3,4,5]. The existing surgical tactics are rather contradictory. Some authors consider it possible to perform primary repair operations, while others are opposed to this approach [6]. However, the experience of domestic and foreign authors shows that the risks and complications that are usually associated with reconstructive surgery may be exaggerated. Neverthe-less, the question of safety and feasibility of pancreatic resection requires further study. The difficulty and uncertainty of complete hemostasis is still one of the main factors determining the restrained attitude of surgeons to resection.

P U R P O S E O F T H E W O R KThe management of pancreatic trauma is com-

plex. The purpose is to report our experience in the management of pancreatic trauma with consideration of morbidity and mortality.


https://dx.doi.org/10.35630/2199-885X/2021/11/2/S.1

M A T E R I A L S A N D M E T H O D SAll patients hospitalized between 1998 and 2017

for pancreatic trauma in the clinic of the Department of Surgical Diseases of the Faculty of Dentistry of Astrakhan State Medical University were investigated. Traumatic injuries of the pancreas were classified ac-cording to the American Association for Surgery of Trauma (AAST) in five grades [3]. We observed 12 patients with pancreatic injuries. There were only male patients among them. They were distributed according their age as follows (Table 1).

According to the nature of injuries, half of the patients were with gunshot injuries, 4 — with blunt injuries (Table 2).

All injuries were combined, the localization of wounds and their size are presented in Table 3.

The size of the wounds from 1 to 2 cm prevailed among all the patients and amounted to 5 (42%) people. The most frequent lesion was to the pancreatic body — 6 (50%) patients.

R E S U L T S A N D D I S C U S S I O NIn seven patients, the wounds were sutured with

interrupted stitches, in one case there was bleeding, as a result of which relaparotomy was performed. The bleeding was stopped by suturing the gland tissue. In one patient, in addition to the nodal suture, the peri-toneum was used as a lining material, and in another, a strand of the omentum on the vascular stem was used. In one case, the gland wound was sutured with an 8-shaped suture. In one patient a resection of the pancreatic tail with suturing of the resulting U-shaped suture wound was performed. In one patient, we used a gastric graft on the vascular stem. When forming a gastric graft, a serous-muscular flap with the right gastroepiploic artery was cut out from the greater curvature of the stomach. This case is of a particular interest, so it is worth being presented.

Patient P. 17 years old, IB No. 3743, was admitted to the clinic with diffuse peritonitis a day after being injured in a motor vehicle accident. The patient under-went laparoscopy, during which a sanious liquid with flakes and jelly-like clots was found in the abdominal cavity. Midline laparotomy was performed. There was a moderate amount of sero-hemorrhagic exudate in the abdominal cavity. There were multiple stearic plaques

S U R G E R Y

https://orcid.org/0000-0002-4303-3764

https://orcid.org/0000-0002-5722-1117

https://orcid.org/0000-0002-7803-8661

https://dx.doi.org/10.35630/2199-885X/2021/11/2/S.1


Table 1. Distribution of patients by age

Table 2. The distribution of the patients according to the nature of injuries

Table 4. Characteristics of patients who underwent laparotomy

males In totalUnder 20 years old 3 (25%) 3 (25%)From 21 to 30 years old 4 (33,3%) 4 (33,3%)From 31 to 40 yaers old 5 (42%) 5 (42%)In total 12 12

The nature of injuries

Blunt trauma

Stab and slash wound

Gunshot wounds In total

males 4 (33,3%) 2 (17%) 6 (50%) 12

Table 3. Localization and size of wounds

Localization of wounds Up to 1 cm From 1 to 2 cm From 2 to 3 cm 8 cm The whole surface In totalThe whole pancreas 1 (8,3%) 1 (8,3%)The body of the pancreas 1 (8,3%) 2 (17%) 3 (25%) 6 (50%)The head of the pancreas 3 (25%) 3 (25%)The tail of the pancreas 1 (8,3%) 1(8,3%) In total 1 (8,3%) 5 (42%) 4 (333%) 1 (8,3%) 1 (8,3%) 12

Patient (n) Mechanism of Injury AAST grade Surgical management Postoperative complication1 Penetrating trauma 3 Distal pancreatectomy None2 Motor vehicle accident 3 Suturing the wound+ gastric graft on the vascular stem None3 Penetrating trauma 1 Suturing the wound Pancreatic fistula4 Motor vehicle accident 1 Suturing the wound None5 Motor vehicle accident 2 Suturing the wound acute pancreatitis6 Motor vehicle accident 2 Suturing the wound None7 gunshot wound 2 Suturing the wound peritonitis8 gunshot wound 2 Suturing the wound None9 gunshot wound 2 Drainage acute pancreatitis10 gunshot wound 2 Drainage None11 gunshot wound 2 Drainage peritonitis12 gunshot wound 2 Drainage intra-abdominal bleeding

on the large and small omentum. The omentum sac was opened, and a large amount of cloudy effusion was released. The revision revealed a large retroperitoneal hematoma extending to the small omentum and me-senteric root. After opening the posterior peritoneum above the pancreas, a complete transverse rupture of its body was found at the borders of the head and body.

The gland was hyperemic with areas of necrosis at the site of rupture. An attempt to isolate the Wirsung duct failed due to rapid infiltration of glandular tissues and of uprisal bleeding. The bleeding was stopped. Necrotic tissues were removed and the wound surface of the distal part of the pancreas was covered with a gastric graft. After the mobilization of the pancreatic body, it was possible to detect the Wirsung duct with a diameter of up to 0.8 cm. A chlorvinyl tube was inserted into its lumen, which was removed in the left hypochondrium. Interrupted stitches were applied to the proximal part. A month after the discharge, the patient developed clinical signs of acute adhesive intestinal obstruction. Relaparotomy was performed. During the revision, the cause of the obstruction was a tube that drained the duct, and good engraftment of

the gastric graft was noted in the area of the previ-ous operation. The gastric graft had a normal color, without signs of swelling. Adhesions were dissected, intestinal obstruction was eliminated. It was decided to transfer external drainage to the internal one. Pancrea-ticogastrostomy was performed. The patient made an uneventful postoperative recovery.

S U R G E R Y


In this case, we observed good engraftment of the gastric graft against the background of existing diffuse peritonitis, that is, in conditions of infection of the abdominal cavity. The distal part of the gland was sutured with interrupted stitches. Organ-preserving surgery was also performed in order to avoid unneces-sary splenectomy and possible violation of the endo-crine function of the pancreas.

During repeated surgery for intestinal obstruc-tion, the external pancreatic fistula was transferred to the internal one, not only because of the high degree of physiology of this method of drainage, but also for reasons of preventing repeated adhesive intestinal obstruction.

The postoperative period was difficult in 6 (50%) patients: 2 (17%) of them had acute pancreatitis; 1 (8,3%) pancreatic fistula; 2 (17%) diffuse peritonitis; 1 (8,3%) bleeding.

The findings reveal two lethal cases among our patients. In the first patient there were acute post-traumatic pancreatitis, pancreatic necrosis, diffuse purulent-fibrinous peritonitis, multiple organ failure and intoxication. The second patient had a complete rupture of the pancreatic body. He developed diffuse purulent-fibrinous peritonitis, multiple organ failure, and cerebral oedema.

C O N C L U S I O N Operative management of pancreatic trauma may

lead to a higher mortality. This must not be necessar-ily related to the pancreas injury alone but due to the associated injuries such as liver, spleen and vascular traumas are more likely to occur with severe outcomes.

As stated above, pancreatic injuries are quite rare. Nevertheless, they are of particular interest, as they may develop severe and sometimes even fatal compli-cations. The gastric graft acts as a barrier to prevent the leakage of pancreatic fluid. It promotes hemostasis, due to the improvement of vascularization and nutri-tion of pancreatic tissues. Moreover, the gastric graft contributes to a more rapid subsidence of pancreatitis, whose manifestations are clinically shown. It should be noted that the postoperating period was unevent-ful and it required less hospital stay after the surgery. The presented data and the methods of performing a gastric graft on the vascular stem in cases of pancreatic damage can be recommended for clinical practice.

R E F E R E N C E S1. Du B., Wang X., Wang X., Shah S., Ke N. A rare

case of central pancreatectomy for isolated complete pancreatic neck transection trauma. BMC Surg. 2019; 19: 91. doi: 10.1186/s12893-019-0557-x

2. Iacono C., Zicari M., Conci S., Valdegamberi A., De Angelis M., Pedrazzani C., Ruzzenente A., Guglielmi A. Management of pancreatic trau-ma: A pancreatic surgeon's point of view. Pancreatol-ogy. 2016;16(3):302-8. doi: 10.1016/j.pan.2015.12.004.

3. Menahem B., Lim C., Lahat E., Salloum C., Os-seis M., Lacaze L., Compagnon P., Pascal G., Azoulay D. Conservative and surgical management of pancreatic trauma in adult patients. Hepatobiliary Surg Nutr. 2016 Dec; 5(6): 470–477. doi: 10.21037/hbsn.2016.07.01

4. Patrizio Petrone, Sara Moral Álvarez, Marta González Pérez, José Ceballos Esparragón, Corrado P Marini. Pancre-atic trauma: Management and literature review Cir Esp. 2017;95(3):123–130. doi: 10.1016/j.cire-sp.2016.05.011.

5. Serra F., Barbato G., Tazzioli G., Gelmini R. Pancreaticogastrostomy as reconstruction choice in pancreatic trauma surgery: Case report and review of the literature. Int J Surg Case Rep. 2019;65:102–106. doi: 10.1016/j.ijscr.2019.10.030

6. Siboni S., Kwon E., Benjamin E., Inaba K., Demetriades D. Isolated blunt pancreatic trauma: A benign injury? J Trauma Acute Care Surg. 2016 Nov; 81(5):855–859. doi: 10.1097/TA.0000000000001224

S U R G E R Y


COMPLICATIONS OF OPEN SURGICAL PROCEDURES IN PATIENTS WITH PANCREATIC CYSTS

A B S T R A C T — The incidence of pancreatic cysts is constantly growing due to an increasing number of patients with a history of pancreatitis. Furthermore, complications after surgical interventions for pancreatic cysts occur in 30-40% of patients. Objective: to analyze the complications in patients with pancreatic cysts after open surgery.We observed 68 patients who underwent open surgical interventions: cystogastrostomy and cystojejunostomy. All patients with pancreatic cysts were divided into 2 groups depending on the type of procedure. In 27 patients (29%) of the first group cystogastrostomy and in 41 patients (81%) of the second group cystojejunostomy was performed.In 1st group complications were diagnosed in 8 cases (29.6%): postoperative pancreatitis (5 — 18.5%), cyst recurrence (3 — 11.1%). In the second group complications occurred in 7 patients (17.1%): postoperative pancreatitis in 4 cases (9.8%), recurrent cysts in 3 cases (7.3%).Thus, cystojejunostomy is less associated with complications than cystogastrostomy.

K E Y W O R D S — pancreatic cyst, chronic pancreatitis, cystogastrostomy, cystojejunostomy.

Aleksandr Gerasimov , Valeriy Nikolskiy ,Aleksandr Mitroshin , Sergey Frolov ,Ekaterina Titova

Penza State University, Medical Institute, Penza, Russia

[email protected]

R E L E V A N C EAcute pancreatitis is considered the single most

frequent gastrointestinal cause of hospital admissions in the United States. Chronic pancreatitis being lower in incidence noticeably worsens patients’ quality of life. In recent decades researchers worldwide have wit-nessed an increase in patients with a history of pancrea-titis, and as a result, the presence of pancreatic cysts in this group of patients [1, 2, 3]. According to scientific literature the process of formation of pancreatic cysts is often accompanied by the appearance of severe complications: mechanical jaundice, perforation, suppuration of cysts, peritonitis, bleeding, etc. [4, 5, 6]. After open surgical interventions complications occur in 30–40% of cases. Those complications are quite variable: from the most severe, such as bleeding, peri-

tonitis, abscesses, to the milder, that could be stopped conservatively or with the help of minimally invasive procedures — postoperative pancreatitis, recurrent pancreatic cysts. These complications are often associ-ated with high pressure on the anastomosis caused by increased pancreatic secretion, as well as technical intraoperative difficulties [7, 8].

Objective: to evaluate complications in patients with pancreatic cysts after open surgical procedures.

M A T E R I A L A N D M E T H O D SThe work has been carried out in surgical depart-

ments of Penza Regional Clinical Hospital named after N. N. Burdenko. The study involved 68 patients who underwent open surgical interventions: cystogastros-tomy and cystojejunostomy. There were 12 women (20.7%) and 56 men (79.3%). The age of patients ranged from 22 to 74 years. The average age was 57±9.8 years. Pancreatic cysts were identified: in the tail of the gland in 6 patients (10.3%), in the body of the pan-creas in 8 patients (13.7%), in the head of the pancreas in 54 patients (75.7%).

At the first stage 59 patients (84.4%) underwent external drainage of pancreatic cysts under ultrasound control. The remaining 6 patients (15.5%) did not undergo this procedure.

Indications for external drainage of cysts were: an increase in the size of the cyst more than 50 mm, a loose, unformed cyst wall, signs of infection of cysts, as well as the severity of the patient's condition.

For diagnostic purposes in the patients with pan-creatic cysts the following general clinical and labora-tory tests were performed: blood type and Rh factor, common blood test, biochemical blood test, general urine test, urine diastasis, coagulogram; instrumental examinations: electrocardiography, ultrasound exami-nation of the abdominal cavity and retroperitoneal space, fibrogastroduodenoscopy, computed tomog-raphy of the abdominal cavity and retroperitoneal space, X-ray examination of the chest and abdominal cavity (fistulography after drainage of pancreatic cysts was performed to identify the connection with the pancreatic duct), magnetic resonance imaging of the abdominal cavity and retroperitoneal space.

S U R G E R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/13

https://orcid.org/0000-0002-9453-7380

https://orcid.org/0000-0001-5158-9570

https://orcid.org/0000-0002-2232-129X

https://orcid.org/0000-0001-9809-0718

https://orcid.org/0000-0002-7674-2057

http://dx.doi.org/10.35630/2199-885X/2021/11/2/13


All patients with pancreatic cysts were divided into 2 groups, depending on the type of surgery per-formed. Group 1 included 27 patients who underwent cystogastrostomy (29%), group 2 included 41 patients where cystojejunostomy was done (81%).

Indications for cystogastrostomy and cystoje-junostomy were: the size of the pancreatic cyst more than 60 mm, the formation of the pancreatic cyst wall, as well as the presence of a proven instrumental con-nection between the cyst and the pancreatic duct.

Cystrogastrostomy was performed in patients with a pronounced adhesive process in the abdomi-nal cavity, with the fusion of the cyst wall with the posterior wall of the stomach. Cystojejunostomy was performed in all remaining cases.

R E S U L T S A N D D I S C U S S I O N In group 1 after surgery, complications were diag-

nosed in 8 cases (29.6%): cyst recurrence in 3 patients, postoperative pancreatitis in 5 patients.

In 5 cases (18.5%), the development of postopera-tive pancreatitis was revealed with clinical manifesta-tions on the 4th–5th day after surgery. After conserva-tive therapy, patients were discharged in 15–16 days in a satisfactory condition. The following scheme of conservative treatment has been used: non-narcotic analgetics (ketorol 1.0 v/m), NSAIDs (ibuprofen 3.0 v/m), antispasmodics (no-shpa 2% — 2.0, platy-phylline 0.2% — 1.0 v/v), infusion therapy in the volume of 20–30 ml/kg of weight was used; drugs that reduce pancreatic secretion (atropine 0.1%–1.0 i/v), as well as inhibitors of protease activity (kontrikal 20,000 units 2 p.i/v or gordox 100.000 units).

In the long-term postoperative period in 3 cases (11.1%) the cyst recurrence was detected (after 4 months during a routine ultrasound examination). No clinical manifestations of pancreatitis were noticed. The level of amylase activity in all cases was within the normal range. The size of the liquid collector in the first case was 18 mm, in the second — 29 mm, in the third — 38 mm. Dynamic follow-up at the surgeon's place of residence is recommended.

Complications occurred in 7 patients (17.1%) in the 2nd group. Postoperative pancreatitis was ob-served in 4 patients (9.8%). In the course of treatment (conservative therapy) acute pancreatitis was stopped on the 12th–16th day from the moment of hospitaliza-tion. Recurrence of a pancreatic cyst was detected during a routine ultrasound examination in 3 patients (7.3%) in the long-term postoperative period (from 4 to 6 months after surgery). Infection of the cyst cavity occurred in one case. In the postoperative period this patient suffered from acute pancreatitis. During the next hospital readmission, the liquid formation in the

projection of the tail of the pancreas with dimensions of 75×62×67 mm was revealed. The cyst cavity was drained under ultrasound control, and cloudy contents with a fetid smell were obtained. After that procedure no clinic of acute pancreatitis were revealed.

After the surgical treatment in the second group of patients the length of hospital stay was maximum 16 days, minimum — 7 (12 ± 1.4 days). In the first group the period of hospital stay was 15 ±1.35 (from 12 to 19 days).

Thus, the total number of complications in 1 group was 29.6%, where postoperative pancreatitis was detected in 18.5%, and recurrent pancreatic cysts — in 11.1%. In 2 group patients, complications were observed in 17.1%: postoperative pancreatitis (9.8%) and recurrent pancreatic cysts (7.3%). The duration of treatment in patients after cystograstrostomy was 15 days, after cystojejunostomy — 12 days. There were no fatal outcomes after surgical interventions in patients with pancreatic cysts.

C O N C L U S I O NWe consider it more appropriate to perform

Roux-en-Y cystojejunostomy on a disconnected loop, since the postoperative period with such a surgical in-tervention is accompanied by fewer complications (by 1.7 times — p=0.017) and a shorter length of hospital stay. The median postoperative LOS after cystogastros-tomy is 1.25 times more than after cystojejunostomy.

R E F E R E N C E S1. Yadav D., Lowenfels A. B. The Epidemiology of

Pancreatitis and Pancreatic Cancer. Gastroenterology. 2013, 144(6): 1252–1261. https://doi.org/10.1053/j.gastro.2013.01.068

2. Kozhevnikov V.V., Gerasimov A.N., Gerasi-mov N.A., Morozov V.S. The result of treatment of patients with bleeding from postnecrotic cysts of the pancreas // In the collection: National projects: chal-lenges and solutions. Materials of the 55th interregional scientific and practical medical conference. Ministry of Health of the Ulyanovsk Region; GBU SPb Research Institute of Ambulance named after N.I. I.I. Janelidze. Ulyanovsk, 2020. P. 487–488.

3. Beyer G., Goni E., Köpke M., G D'Haese J., Werner J., Schirra J., Mayerle J. Management Algorithm for Cystic Pancreatic Lesions // Visc Med. 2018. Jul;34(3):197–201. doi: 10.1159/000489233.

4. Sklyarenko R.T., Dovgalyuk A.Z., Omarov M.A., Spiridonova V.S., Shabanova O.A. Consequences of surgical treatment of diseases of the pancreas (chronic pancreatitis, cysts and fistulas of the pancreas) // In the book: Clinical and expert charac-teristics of persistent dysfunctions in surgical diseases and the possibility of rehabilitation. St. Petersburg. – 2017. P. 355–375.

S U R G E R Y


5. Cherdantsev D.V. Diagnostics of postnecrotic pancreatic cysts (literature review) / Cherdantsev D.V., Pervova O.V., Noskov I.G., Fokin D.V., Kazadaeva A.A., Kazadaeva I.A. // Messenger of Surgery I.I. Grekov. – 2020. Vol. 179. N. 2. P. 68–72.

6. Koshel A.P., Drozdov E.S., Dibina T.V., Klok-ov S.S., Mironova E.B., Rakina Yu.Yu. Combined method for differential diagnosis of cystic neoplasms of the pancreas // Siberian Journal of Oncology. 2018. – Vol. 17. – N.6. – P. 27–34.

7. Chudoba A, Degowska M, Oracz G. Multilocu-lar Cyst of the Pancreas // Indian J Pediatr. 2020. N. 87(11). P. 965.

8. Cao C.X., Sharib J.M., Blanco A.M., Gold-berg D., Bracci P., Mukhtar R.A., Esser-man L.J., Kirkwood K.S. Abdominal Imaging of Pancreatic Cysts and Cyst-Associated Pancreatic Cancer in BRCA1/2 Mutation Carriers: A Retro-spective Cross-Sectional Study // J Am Coll Surg. 2020 Jan;230(1):53–63.e1. doi: 10.1016/j.jamcoll-surg.2019.09.019.

S U R G E R Y


LITERATURE REVIEWMETHODS FOR PROSTHETIC MESH FIXATION IN SURGICAL REPAIR OF INGUINAL HERNIAS

A B S T R A C T — The article is devoted to the evolution of fixation methods for synthetic mesh endoprostheses in surgery of inguinal hernias. We have analyzed the data on failures and complications of synthetic implants. We paid a special attention to available techniques, such as endoprostheses fixation without sutures as well as to the fixation using various types of glue. Our review is based on its own experimental and clinical data as well as the data from world's best hospitals. We found that the use of fibrin glue frequently leads to formation of seromas and hematomas, whereas albumin-glutaraldehyde glue may result in abscesses and pyogenic inflammatory infections of surgical sites. The authors pointed out the importance of further research for the optimal fixation of the prosthetic mesh in hernia repair.

K E Y W O R D S — inguinal hernias, hernia prosthesis, mesh implants, methods of fixation, postoperative complications, surgical glue, surgical hernia repair.

Vladimir Akimov1 , Dmitry Krikunov1 ,Dmitry Parshin2 , Behruz Radzhabov1 ,Vladimir Senko3

1 I.I. Mechnikov North-Western State Medical University, Saint Petersburg;2 Astrakhan State Medical University, Astrakhan;3 L.G. Sokolov Clinical Hospital No 122, Saint Petersburg, Russia

[email protected]

At the end of the twentieth century, a lot of stud-ies were published, where the analysis of the causes of recurrence after hernia repair was carried out. The main reason for their development was the migration of the implant, so the need for secure fixation of the mesh was reported [3]. In the literature, there are sev-eral reasons for the migration of implants — mechani-cal (primary) and those caused by dystrophic changes in the tissues surrounding the implant (secondary) [4]. Primary ones arise mainly because of inadequate fixa-tion of the mesh implant [4].

In terms of modern hernia repair technologies, many questions come up about the possibilities of sutureless fixation and their advantages compared to traumatic (penetrating) fixation. The method of sutureless fixation was introduced into clinical practice of premises of our department in 2015; however, the ex-perimental study of this technique had been conducted in the United States and European countries since the beginning of the second decade of the 21st century and has been continued up to the present day [5]. The pos-sibilities of body tissues connection still causes much discussion, and the scientific search is going on [6].

Chronic pain after hernioplasty occurs in addition to many external factors, from the body's inflammatory response to the implant and the involvement of sensory nerve endings up to a psychoemotional state of the pa-tient. However, the leading cause of neuralgia, pain, and paresthesia after inguinal hernia repair is the involvement of sensory nerve endings in the suture area or in inflam-matory tissues after placement of hernia implant [7].

M E T H O D SWe selected the published data, including

PubMed and Web of Science databases on the in-fluence of the method of fixing mesh prostheses in the surgical treatment of hernias. 118 sources were analyzed from 2003 to 2020, and 27 sources included in the article.

R E S U L T S O F S E L E C T I O N A N D C O N T E N T O F T H E R E V I E W

Among all the methods of sutureless fixation, ParieteneProGrip implant (Sofradim, France) can be

Received 13.03.2021 Received in revised form 30 April 2021; Accepted 7 Mai 2021

I N T R O D U C T I O NIn the conditions of modern inguinal hernia

surgery, there is a tendency to focus on the methods of surgical interventions and types of mesh implants without considering the multiple disadvantages of conventional methods for mesh fixation. Dozens of ex-periments carried out over the past 20 years proved the existence of a pronounced inflammatory process along the periphery of implanted prosthesis (Graft-versus-Host disease), which onset can reach 5–10 years after hernioplasty [1]. As a result, there are many complica-tions emerging after inguinal hernia repair (acute and chronic pain, sense of discomfort, paresthesia, neural-gia, up to the inflammation of the implants and their failure) [2].

S U R G E R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/14

https://orcid.org/0000-0002-7388-0366

https://orcid.org/0000-0002-0242-6895

https://orcid.org/0000-0002-1050-7716

https://orcid.org/0000-0002-7586-6859

https://orcid.org/0000-0002-7813-9673

http://dx.doi.org/10.35630/2199-885X/2021/11/2/14


pointed out. It consists of a mesh with micro-grips represented by polypropylene fibres and resorbable polylactic acid micro hooks on its lower surface, thanks to which it is fixed (“Velcro”) [8]. The results of the first experience of using this type of mesh in the Russia by Protasov A.B. are very positive: the number of complications decreased by sixfold and duration of the operation declined by 1.4 times [9].

There is also a number of rigid meshes with shape-memory features, for example Hertramesh (Herniamesh, Italy), which, according to the authors, do not require any fixation. However, during the stud-ies the authors noticed significantly more frequent de-velopment of various kinds of complications (chronic pain, foreign body sensation, neuralgia and paresthe-sia) in comparison to lightweight meshes [10].

The use of adhesive compositions for mesh fixation during the inguinal hernia surgical repair can be particularly highlighted [11]. Before the start of our first publications (Chair of Surgery named after N.D. Monastyrskiy) on the use of adhesive composi-tions [12, 13] there were few publications on this issue in the Russia, and most surgeons had no idea about the use of this method of fixation. With the beginning of development and the first demonstrations, several dozen publications appeared dealing with the use of various kinds of glue mesh fixation methods.It con-firms the value and need for further development of this method of fixation in our country. According to the foreign literature data, the inflammatory process in the mesh area after the use of adhesive compositions is much less obvious at the edges [14, 15]. In the modern world, there are various types of glue used for inguinal hernia repair; the most common is Fibrin glue that is used in traditional methods of inguinal hernia repair. However, there are reports of the possibility of its use in endovideosurgical methods of hernioplasty [16].

In 2014, The European Hernia Society did not include direct instructions on the use of adhesive compositions for mesh fixation in the EHS Guide-lines for the Treatment of Inguinal Hernia, but it was mentioned that this technology can lead to a decrease in the risk of chronic pain development, but there is not enough studies on this subject [17].

The use of cyanoacrylates for inguinal hernia surgical repair is of great interest [21]. In 2007 Aus-trian authors carried out an experiment demonstrating the undesirability of using cyanoacrylates in hernio-plasty, as, in their opinion, the glue was not resorbed and it caused persistent deformation of the anterior abdominal wall in the area of its application. In the next experiment that was carried out more than 5 years later, the complete safety of cyanoacrylate use and their complete resorption after 40–50 days from

application was proved; the authors associate the unsatisfactory results of the Austrian colleagues with applying too much glue [22]. Up to date, two works have been published concerning the use of cyanoacr-ylate glue (Switzerland, Finland, 2011). The first study compares the use of lightweight meshes and cyanoacr-ylate glue in traditional hernioplasty with the same lightweight meshes and absorbable sutures. The results indicated 2 times higher percentage of recurrences but at the same time 2 times lower level of chronic pain while using the glue fixation method [23]. The second study, published 5 years later, showed an identical level of recurrence and chronic pain development; however, the authors noted the presence of wound suppuration while using glue fixation — in 3.4% of cases.

C O N C L U S I O NBased on the experiment data, it can be con-

cluded that:— the use of traumatic fixation methods (for

example, sutures), reduces mesh contraction in the postoperative period [18];

— the use of fibrin glue is associated with fewer postoperative complications such as seromas or he-matomas [19];

— the use of albumin-glutaraldehyde glue causes formations abscess and other pyoinflammatory dis-eases in the prosthetic mesh area [20].

Considering the above, it is rather difficult to es-timate the possibility and safety of using glue fixation methods for hernia repair; this issue requires further study and a series of experiments to make any conclu-sions [24]. Of course, it is necessary to conduct further research on this issue and major clinical experimental studies [25].

R E F E R E N C E S1. Aiolfi A, Cavalli M, Micheletto G, Lom-

bardo F, Bonitta G, Morlacchi A, Bruni PG, Campanelli G, Bona D. Primary inguinal hernia: systematic review and Bayesian network meta-analysis comparing open, laparoscopic transabdominalpreperi-toneal, totally extraperitoneal, and robotic preperi-toneal repair. Hernia. 2019;23(3):473–484. DOI: 10.1007/s10029-019-01964-2.

2. Bittner R, Bain K, Bansal VK, Berrevoet F, Bingener-Casey J, Chen D, Chen J, Chowbey P, Dietz UA, de Beaux A, Ferzli G, Fortelny R, Hoffmann H, Iskander M, Ji Z, Jorgensen LN, Khullar R, Kirchhoff P, Köckerling F, Kukleta J, LeBlanc K, Li J, Lomanto D, Mayer F, Meytes V, Misra M, Morales-Conde S, Niebuhr H, Radvinsky D, Ramshaw B, Ranev D, Reinpold W, Sharma A, Schrittwieser R, Stechemesser B, Sutedja B, Tang J, War-ren J, Weyhe D, Wiegering A, Woeste G, Yao Q. Update of Guidelines for laparoscopic treatment

S U R G E R Y


of ventral and incisional abdominal wall hernias (International Endohernia Society (IEHS))-Part A. SurgEndosc. 2019;33(10):3069–3139. DOI: 10.1007/s00464-019-06907-7.

3. Jenkins ED, Melman L, Desai S, Brown SR, Frisella MM, Deeken CR, Matthews BD. Evaluation of intraperitoneal placement of absorbable and nonabsorbable barrier coated mesh secured with fibrin sealant in a New Zealand white rabbit model. SurgEndosc. 2011;25(2):604–12. DOI: 10.1007/s00464-010-1230-8

4. Ielpo B, Duran H, Diaz E, Fabra I, Caruso R, Malavé L, Ferri V, Lazzaro S, Kalivaci D, Quijano Y, Vicente E. A prospective randomized study comparing laparoscopic transabdominalpreperi-toneal (TAPP) versus Lichtenstein repair for bilateral inguinal hernias. Am J Surg. 2018;216(1):78–83. DOI: 10.1016/j.amjsurg.2017.07.016.

5. Raakow J, Aydin M, Kilian M, Köhler A, Werner S, Pratschke J, Fikatas P. Elektive Versorgung von Leistenhernien in der universitären Chirurgie – eine ökonomische Herausforderung [Elective treatment of inguinal hernia in university sur-gery-an economic challenge]. Chirurg. 2019;90(12): 1011–1018. DOI: 10.1007/s00104-019-1008-z.

6. Bittner R, Montgomery MA, Arregui E, et al. Update of guidelines on laparoscopic (TAPP) and endoscopic (TEP) treatment of inguinal hernia (International Endohernia Society) [published cor-rection appears in SurgEndosc. 2015;29(6):1655–6. DOI:10.1007/s00464-014-3917-8.

7. Topchiev M, Parshin D, Nurmagomedov A, Topchiev A, Mukhtarov I. Tensometric charac-teristics of de-epithelized skin graft and polypropylene mesh implant used in hernioplasty. ArchivEuro-medica. 2019;9(3):55–57. DOI: 10.35630/2199-885X/2019/9/3.18.

8. Bittner R, Arregui ME, Bisgaard T, et al. Guidelines for laparoscopic (TAPP) and endo-scopic (TEP) treatment of inguinal hernia [Inter-national Endohernia Society (IEHS)]. SurgEndosc. 2011;25(9):2773–2843. DOI:10.1007/s00464-011-1799-6.

9. Bullen NL, Massey LH, Antoniou SA, Smart NJ, Fortelny RH. Open versus laparoscopic mesh repair of primary unilateral uncomplicated inguinal hernia: a systematic review with meta-analysis and trial sequential analysis. Hernia. 2019;23(3):461–472. DOI: 10.1007/s10029-019-01989-7.

10. Chai FY. The "Best" Inguinal Hernia Repair Tech-nique. World J Surg. 2020;44(10):3322–3323. DOI: 10.1007/s00268-020-05636-6.

11. Protasov A, Topchiev A, Parshin D, Brus-nev L, Emkuzhev K, Mukhtarov, I. Analysis of postoperative complications in repair of incisional ventral hernias using allo- and autografts. ArchivEuro-medica. 2020;10(4):107–109. DOI: 10.35630/2199-885X/2020/10/4.26.

12. Gavigan T, Stewart T, Matthews B, Reinke C. Patients Undergoing Parastomal Hernia Repair Using the Americas Hernia Society Quality Collabo-rative: A Prospective Cohort Study. J Am Coll Surg. 2018;227(4):393–403. DOI: 10.1016/j.jamcoll-surg.2018.07.658.

13. Ferrarese A, Bindi M, Rivelli M, Solej M, En-rico S, Martino V. Self-gripping mesh versus fibrin glue fixation in laparoscopic inguinal hernia repair: a randomized prospective clinical trial in young and eld-erly patients. Open Med (Wars). 2016;11(1):497–508. DOI: 10.1515/med-2016-0087.

14. Akimov V.P., KrikunovD.Yu., Toidze V.V., Churgulia M.Z., Kashchenko V.A. Pos-sibilities of using a cyanacryl adhesive for fixation of a net implant in the laparoscopic treatment of animal herbs. Vestnik of Saint Petersburg Univer-sity. 2018;13(3):282–290. DOI: 10.21638/11701/spbu11.2018.305.

15. KrikunovD, Akimov V, Toidze V, Churgulia M. Comparative evaluation of TAPP hernioplasty with use of various methods of fixing the reticular endoprosthesis and TEP in the treat-ment of inguinal hernias. Georgian Medical News. 2018;(278):15–20. PMID: 29905538.

16. Bansal VK, Misra MC, Babu D, Victor J, Kumar S, Sagar R, Rajeshwari S, Krishna A, Rewari V. A prospective, randomized comparison of long-term outcomes: chronic groin pain and quality of life following totally extraperitoneal (TEP) and transabdominalpreperitoneal (TAPP) laparoscopic inguinal hernia repair. Surg Endosc. 2013;27(7): 2373–82. DOI: 10.1007/s00464-013-2797-7.

17. de Goede B, Wijsmuller AR, van Ramshorst GH, van Kempen BJ, Hop WCJ, Klitsie PJ, Scheltinga MR, de Haan J, Mastboom WJB, van der Harst E, Simons MP, Kleinrensink GJ, Jeekel J, Lange JF; INCA Trialists’ Collaboration. Watchful Waiting Versus Surgery of Mildly Symp-tomatic or Asymptomatic Inguinal Hernia in Men Aged 50 Years and Older: A Randomized Controlled Trial. Ann Surg. 2018;267(1):42–49. DOI: 10.1097/SLA.0000000000002243.

18. Bittner R, Schwarz J. Inguinal hernia repair: current surgical techniques. Langenbecks Arch Surg. 2012;397(2):271–82. DOI: 10.1007/s00423-011-0875-7.

19. Miserez M, Peeters E, Aufenacker T, Bouil-lot JL, Campanelli G, Conze J, Fortelny R, Heikkinen T, Jorgensen LN, Kukleta J, Morales-Conde S, Nordin P, Schumpelick V, Smedberg S, Smietanski M, Weber G, Simons MP. Update with level 1 studies of the European Hernia Society guidelines on the treatment of inguinal hernia in adult patients. Hernia. 2014;18(2):151–63. DOI: 10.1007/s10029-014-1236-6.

20. Ahmad M, Niaz WA, Hussain A, Saeeduddin A. Polypropylene mesh repair of incisional hernia. J Coll Physicians Surg Pak. 2003;13(8):440–2. PMID: 12921679.

S U R G E R Y


21. Chen DC, Morrison J. State of the art: open mesh-based inguinal hernia repair. Hernia. 2019;23(3):485–492. DOI: 10.1007/s10029-019-01983-z.

22. Ashrafi D, Siddaiah-Subramanya M, Memon B, Memonm.A. Causes of recurrences after open inguinal herniorrhaphy. Hernia. 2019;23(4):637–645. DOI: 10.1007/s10029-018-1868-z.

23. Schmidt L, Öberg S, Andresen K, Rosenberg J. Laparoscopic repair is superior to open techniques when treating primary groin hernias in women: a nationwide register-based cohort study. Surg Endosc. 2019;33(1):71–78. DOI: 10.1007/s00464-018-6270-5. Epub 2018 Jun 15. PMID: 29905895.

24. Tastaldi L, Barros PHF, Krpata DM, Prabhu AS, Rosenblatt S, Petro CC, Alkhatib H, Szutan LA, Silva RA, Olson MA, Stewart TG, Roll S, Rosen MJ, Poulose BK. Hernia recur-

rence inventory: inguinal hernia recurrence can be accurately assessed using patient-reported outcomes. Hernia. 2020;24(1):127–135. DOI: 10.1007/s10029-019-02000-z.

25. Aiolfi A, Cavalli M, Bonitta G, Bona D. Inguinal hernia repair in centers of excellence. Hernia. 2020;24 (1):215. DOI: 10.1007/s10029-019-02010-x.

26. AlMarzooqi R, Tish S, Huang LC, Prabhu A, Rosen M. Review of inguinal hernia repair techniques within the Americas Hernia Society Quality Collabo-rative. Hernia. 2019;23(3):429–438. DOI: 10.1007/s10029-019-01968-y.

27. Drs A, Horák P, Chlupáč J, Froněk J. The most recent recommendations for the surgical treatment of inguinal hernia. RozhlChir. 2019;98(7):268–272.PMID: 31398985.

S U R G E R Y


EXPERIENCE OF ENDOVASCULAR EMBOLIZATION FOR RECTAL BLEEDING IN PATIENTS WITH RECTAL CANCER

A B S T R A C T — The article is devoted to the most dangerous complication of colorectal cancer — bleeding. The clinical group included 32 patients with stages 3–4 of the disease. Endovascular embolization by microspheres was performed in 18 patients. The bleeding was successfully stopped in all operated patients. When monitoring the main laboratory blood values in the postoperative period, the development of a systemic inflammatory reaction was not noted. Flexible rectosigmoscopy performed after endovascular embolization did not reveal a single case of necrosis of the intestinal wall above and below the tumor. In order to prevent intestinal ischemia, it is advisable to introduce microspheres in a volume of ⅓ from the circulatory channel of the embolized area. Endovascular embolization by microspheres is an effective method of hemostasis in patients with colorectal cancer complicated by bleeding.

K E Y W O R D S — colorectal cancer, rectal cancer, rectal bleeding, endovascular embolization, endovascular hemostasis, microspheres.

Alexander Khitaryan1, Alexander Murlychev1 ,Alexey Orekhov1, Sergey Kovalev1, Vyacheslav Mykhaylichenko2 , Dmitry Parshin3

1 RZD-Medicine Clinical Hospital, Rostov-on-Don, Russia2 S.I. Georgievsky Medical Academy, Simferopol3 Astrakhan State Medical University, Astrakhan, Russia

[email protected]

Received 02 March 2021 Received in revised form 8 May 2021; Accepted 11 May 2021

I N T R O D U C T I O NColorectal cancer (CRC) is the second most

common cause of cancer death in the United States. The incidence rate between 2012 and 2016 ranged from 30 to 45.7 per 100,000 people. Among persons younger than 50 years old, the incidence of tumors of the proximal and distal colon, as well as the rectum, increased by about 2% annually. Given the increasing incidence of CRC in younger patients, in 2018, the American Cancer Society made a qualified recom-mendation to reduce the age at which patients should begin CRC screening to 45 years and older. It was ex-trapolated that in 2020, approximately 147,950 people will be diagnosed with CRC in the US, and 53,200 will die from the disease, including 17,930 cases and 3,640 deaths among persons under the age of 50 [1].

Intestinal bleeding, as a complication of CRC, was noted in 26.8% of patients. The complication is usually associated with the breakdown of tumor tissue or germination and arrosion of intestinal wall vessels. In 21.6% of the total number of observations of occult gastrointestinal bleeding, their source is localized in the rectum, in 11% of the total number of observa-tions, bleeding is associated with carcinoma. Profuse bleeding occurs in 2–5% of all cases of rectal cancer (RC) [2, 3, 4].

RC complicated with continued intense bleeding and pronounced anemia poses challenges for doctors of general surgical practice in preparing the patient for surgery, often with low efficiency of blood-substituting measures. Under certain circumstances you have to refuse to perform neoadjuvant exposure. Postoperative lethality and the frequency of complications in pa-tients with RC operated at the bleeding level is higher than in those operated as planned. In primary resec-tions with anastomosis for complicated RC, lethality varies from 4.3 to 8.9%, the frequency of postoperative complications varies from 21.7 to 89%. The five-year survival rate in these patients is 60.9% versus 74.6% in those operated in the absence of complications [5, 6].

The purpose of the work was to summarize the experience of endovascular embolization in 18 patients with RC admitted to a general surgical hospital with bleeding.

M A T E R I A L A N D M E T H O D SIn clinical group there were 32 patients with

RC T3-4NxM1. In our studies, we adhered to the 7th revision of the TNM tumor classification of the International Cancer Union. 16 (50.0%) men and 16 (50%) women. The age of patients ranged from 60 to 73 years, averaging 65.8 ± 3.1 years. The inclusion cri-teria were: the presence of a powerful collateral from the right branch of the upper rectangular artery to the lower sigmoid artery during angiography. Exclusion criteria were: signs of intestinal obstruction, history of other oncological diseases, pronounced aortic atherosclerosis, concomitant blood diseases (throm-bophilia), sub- and decompensation of diseases of the cardiovascular, respiratory system, liver and urinary system.

S U R G E R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/15

https://orcid.org/0000-0003-0808-1338

https://orcid.org/0000-0003-4204-5912

https://orcid.org/0000-0002-1050-7716

http://dx.doi.org/10.35630/2199-885X/2021/11/2/15


At the first stage of treatment, hemostatic therapy was carried out for 2 hours. With continuing bleeding, a decision was made to conduct diagnostic aortography of the abdominal aorta to identify the source of bleeding. In 12 patients, contraindications to endovascular artery embolization were identified due to the features of angioarchitectonics: pronounced atherosclerotic lesion with aortic calcinosis at the site of withdrawal of the inferior mesenteric artery in 5 patients, pronounced atherosclerotic lesion of the infe-rior mesenteric artery with calcinosis from the mouth in 3 patients, inflection of the mouth of the lower me-senteric artery in one patient which could lead to dis-section of the vessel when conducting the catheter to its distal sections, an additional vessel connecting the rectal artery basin and the sigmoid arterial arcade bear-ing the danger of non-target embolization of healthy organ vessels in 3 patients. Two patients refused to per-form an endovascular intervention. In these patients, bleeding stop continued to be carried out by con-servative means. Embolization was performed on the Philips Allura CV 20 angiographic unit. Femoral 5-6Fr introducers were used for vascular access. Selective catheterization of the inferior mesenteric and superior rectangular arteries was performed by a 4-5F catheter. As emboli, microspheres of HepaSphere of Biosphere Medical (France) were used. The choice of emboles is due to the possibility of their saturation with a cyto-static solution, which in our opinion is promising with the further development of the technique.

The volume of embolisate administered was determined according to the method we developed and was ⅓ of the volume of the arterial circulator bed in the area of selective installation of the catheter for embolization. After embolization, arrest of rectal bleeding was monitored clinically and laboratory.Flexiblerectosigmoscopy was performed in dynam-ics.Attentionwas paid to the presence of hemostasis, superficial necrosis of the tumor, and the condition of the rectal mucosa in the immediate proximity of the tumor. After pre-surgical preparation, all patients were referred for specialized combined RC treatment.

R E S U L T S A N D D I S C U S S I O NTreatment results were evaluated by the quality

of hemostasis, systemic inflammatory response and rectal mucosa conditions above and below the tumor. Hemostasis was achieved in 100% of cases in all 18 pa-tients. According to the results of flexible rectosigmos-copy, the intestinal mucosa above and below the tumor was unchanged. The blood hemoglobin level in the clinical group was reduced and averaged 74.1±11 g/l. In the largest number of observations, blood he-moglobin was distributed in the range of 70–90 g/l

(61.1%, n = 11). To assess the degree of inflammation in the body, all patients had their leukocyte intoxica-tion index (LII)determined.On the 1 and 2 day after the operation, patients’LII, compared with the initial value, was increased to 1.2 ± 0.08 and 0.99 ± 0.07. In subsequent stages of observation, the LII decreased to normal values. A statistically significant decrease in total protein content in the blood compared to the original value was observed on days 1 and 2 after surgery. The total blood bilirubin in the first day after the operation increased, but remained within normal limits throughout the observed stage. There were no significant changes in the urea content in the blood of patients in clinical groups. By 3 days after the opera-tion, the blood urea level decreased, probably due to infusion therapy. Thus, the study of the dynamics of laboratory indicators in patients after endovascular embolization of the tumor due to continued bleeding did not lead to a systemic inflammatory reaction.

C O N C L U S I O NAccording to the results of the postoperative

observation, it can be concluded that endovascular embolization is an effective method of hemostasis in this category of patients.Theintervention does not lead to complications, does not cause a systemic inflamma-tory reaction, does not complicate the course of the underlying disease in the early postoperative period and is easily tolerated by patients. Performing a flexible rectosigmoscopy after endovascular embolization did not reveal a single case of necrosis of the intestinal wall above and below the tumor. In order to prevent intesti-nal ischemia, it is advisable to introduce microspheres in a volume of ⅓ from the circulatory channel of the embolized area.

Contributors: Alexander Khitaryan — designed the study,

performed the analysis and interpreted the results; Alexander Murlychev — conducted review of the literature and collected the data; Alexey Orekhov — performed the analysis and interpreted the results; Sergey Kovalev — collected the data and performed the calculations; Vyacheslav Mikhailichenko — performed the analysis and interpreted the results; Dmitry Parshin — wrote the manuscript.

R E F E R E N C E S1. Siegel, R.L., Miller, K.D., Goding Sauer,

A., Fedewa, S.A., Butterly, L.F., Anderson, J.C., Cercek, A., Smith, R.A. and Jemal, A. Colorectal cancer statistics, 2020. CA A Cancer J Clin. 2020;70: 145–164. https://doi.org/10.3322/caac.21601.

2. Mathis KL, Pemberton JH. New developments in colorectal surgery. Current Opinion in Gastro-

S U R G E R Y


enterology. 2015;31(1):62–68. DOI: 10.1097/mog.0000000000000138.

3. Saliangas, K., Economou, A., Nikoloudis, N. et al. Treatment of complicated colorectal cancer. Evaluation of the outcome. Tech Coloproctol. 2004; 8:199–201). https://doi.org/10.1007/s10151-004-0156-3//.

4. Pham T, Tran BA, Ooi K, Mykytowycz M, McLaughlin S, Croxford M, Skinner I, Faragher I. Super-Selective Mesenteric Emboliza-tion Provides Effective Control of Lower GI Bleed-ing. Radiol Res Pract. 2017;2017:1074804. doi: 10.1155/2017/1074804. Epub 2017 Jan 22. PMID: 28210507; PMCID: PMC5292126.

5. Rachapalli V, Nagabhushan J. Massive rectal bleeding: empiric embolization of the superior rectal artery. ANZ J Surg. 2018;88(1–2):116. doi: 10.1111/ans.14188. PMID: 29392904.

6. Park S, Kim Y, Shin JH, Yang WJ, Noh SY, Chu HH, Chen CS, Yoon HK. Outcome of Rectal Arte-rial Embolization for Rectal Bleeding in 34 Patients: A Single-Center Retrospective Study over 20 Years. J VascIntervRadiol. 2020;31(4):576–583. doi: 10.1016/j.jvir.2019.05.013. Epub 2019 Sep 14. PMID: 31530489.

S U R G E R Y


PRECLINICAL TRIALS FOR ADVANCED CHITOSAN-BASED COATINGS IN TREATING PURULENT WOUNDS

A B S T R A C T — Wound healing is a complex set of body responses to damaged tissues, which manifests itself through local destructive-inflammatory changes and general reactions. There are numerous coatings used currently in clinical practice to treat wounds. These coatings serve to develop a wet anti-bacterial environment, which is essential to facilitate the healing process. This work focuses on studying multifunctional coatings, which are based on chitosan, a biocompatible polymer featuring wound-healing properties. The distinctive feature to be found in chitosan fibers is their density and the orientation of pores. The coatings within this study were tested on scalped full-layer skin wounds of male Wistar-Kyoto rats and male rabbits. The sample with a dense structure and low biological resorption was found to be effective in performing the function of a framework, as well as in ensuring proper drainage at the affected area. This is important when treating purulent wounds.The experimental sample with a high degree of adhesion and a shorter biodegradation life can be recommended for treating wounds with no purulent-inflammatory issues, for granulating wounds, as well as a drug carrier-matrix. The studied wound coatings have revealed their medical efficiency at the preclinical stage (in vivo). Using wound coatings with specified structural and functional features would allow making a reasonable choice when selecting a coating depending on the stage of wound healing course.

K E Y W O R D S — chitosan; purulent wound; infected wound; wound process; wound coatings; wound treatment; preclinical tests; regenerative medicine; tissue reconstruction.

Denis Ushmarov1 , Sergey Gumenyuk1 ,Alexander Gumenyuk1 , Tatyana Gaivoronskaya1, Timofei Grigoriev2 , Ksenia Lukanina2 ,Timofey Patsaev2 , Dmitry Domenyuk3

1 Kuban State Medical University, Krasnodar; 2 National Research Center "Kurchatov Institute" Moscow;3 State Medical University, Stavropol, Russia

[email protected]

Received 20.03.2021 Received in revised form 11 May 2021; Accepted 18 May 2021

the need to develop highly effective wound-healing medicines with antimicrobial properties [1–3].

The main factors that impede epithelialization and granulation include tissue dystrophy, oxidative damage, wound moisture content imbalance, infec-tions and other complications affecting the area of surgical resection, injury or burn. The development of scar tissue or other structural changes at the wound will reduce the patient’s life quality. Nowadays, there are certain techniques available, which can help facilitate the wound healing process as well as improve the structural and functional properties of the newly developing tissue. Most of such techniques involve wound coatings that differ in their composition and functional features [4–6, 25–27].

The data available in respective literature reveals that purulent-inflammatory processes affecting soft tissues are rated among the most common issues in the entire body of surgical morbidity. The pathologies in question account for 35–45% of all surgical cases, while purulent complications, which affect the post-operative wound area, have been observed in 33–38% of all patients. Moreover, as far as outpatient clinical practice is concerned, soft tissue infections constitute the predominant pathology within the total structure of primary surgical patients [7].

The physical methods, which are used to treat wounds nowadays (vacuum treatment of wounds, ultrasonic cavitation, wound treatment with a pulsat-ing jet, etc.) do add to the positive treatment outcome. However, even with all these advanced methods, the treatment of purulent and infected wounds can never be effective without wound dressing [8].

In modern conditions, the physical and hygienic properties featured by materials based on natural cot-ton no longer meet clinical specialists’ needs. Given that, there has been a wider use of dressings made with advanced technologies and featuring a set of the following properties: inertia in relation to biologi-cal tissues; minimal mechanical trauma potential; impermeability to microorganisms and dust particles; pH control; gas composition and humidity of the environment around the wound. Therefore, one of the major requirements for advanced wound coverings (dressings) implies their maximum action polyvalence [9–11].

I N T R O D U C T I O NOne of the urgent issues faced currently by medi-

cine is the treatment of patients with injuries, burns, and various skin defects (e.g., trophic ulcers, bedsores, infected postoperative wounds, etc.). All this indicates

S U R G E R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/16

https://orcid.org/0000-0002-3665-3906

https://orcid.org/0000-0002-4197-2207

https://orcid.org/0000-0002-2294-0808

https://orcid.org/0000-0001-8197-0188

https://orcid.org/0000-0002-7299-4361

https://orcid.org/0000-0001-8736-8032

https://orcid.org/0000-0003-4022-5020

http://dx.doi.org/10.35630/2199-885X/2021/11/2/16


There are over 470 samples of various dressings and wound coverings available now globally and com-ing in various pharmaceutical forms, whereas the basis for their manufacture includes polymers, collagen, gelatin, cellulose, pectin and many other substances, including various combinations of these materials that feature the respective properties and offer enough pathogenetic grounds to use them to treat wounds [12–14]. This means that one of the promising ways to improve the quality of the treatment offered to patients with infected and purulent wounds implies a scientific and experimental search for new dressings or modifying the already available ones.

Taking into account the above-described require-ments for modern wound coatings, one of the most important properties that such materials should possess is biodegradation. From this stance, we believe chitosan — a derivative of the natural polysaccharide chitin — is to deserve special attention [15–18].

The increased interest taken in it seen as a basis for the production of wound coatings can be explained by its capacity to change its physical and chemical prop-erties depending on changes in the molecular weight and three-dimensional structure. At the same time, the form factor of the product can change from a hydrogel to a frame structure with different degrees of density, swelling and moisture absorption. The specific features inherent in this polymer and the materials made on its basis explain the potential for its use. [19].

Such features include minimal side effects, bio-compatibility, high healing potential, moisture- and air-permeability, high porosity, mechanical stability combined with plasticity, and a predictable biore-sorption period within the body. Chitosan possesses hemostatic, bacteriostatic, and fungistatic proper-ties. There have also been immunomodulating and antitumor effects identified, as well as lack of immu-noreactivity has been proven along with complete elimination out of the body and biostimulation of regeneration [20, 21]. The study has also shown that chitosan films can inhibit significantly the growth of microflora (Staphylococcus, Proteus, Pseudomonas aeruginosa), at the same time accelerating the healing of burns. The universal mechanism, which promotes selective binding of chitosan to sugar receptors on the cell membrane, produces a bacteriostatic effect, which, in turn, stops the infectious process involving the most significant microorganisms [22, 23].

In view of all the properties mentioned above, which are featured by the proposed chitosan-based coatings, we can talk about the holding a research project aiming at developing dressings with a polyva-lent action range.

Aim of Study: To expand the range of advanced multifunc-

tional chitosan-based wound coatings with significant antimicrobial and healing properties; to study in vivo the specific features of the wound process involving the treatment of purulent soft tissue wounds of various origins.

M E T H O D SStudy Design

A multi-stage randomized controlled experimen-tal study involved 225 conventional male rats, weigh-ing 250–300 g each — at Stage 1; 135 individuals of linear Wistar male rats, weighing 250–300 g each — at Stage 2, and 27 male rabbits (Soviet Chinchilla breed), weighing 2,900 (±150) g each — at Stage3 of the study.

To create a model of purulent soft tissue wound in experimental animals, we employed our own tech-nique (Patent for invention RU # 2703709 of August 23, 2018). This comes down to developing a wound featuring specified parameters through inserting a rounded polymer implant into the soft tissues for 6-7 days. Wound infection in animals was modeled through the introduction of Staphylococcus aureus (S. aureus 209P (ATCC6538P)) on foam balls for soft application of liquid and paste-type medicines Pele Tim # 3 (VOCO, Germany). To create infecting, a suspension of an 18–20-hour microorganism culture in a saline solution was used (concentration — 105 microbial bodies per 1 ml). 0.1 ml of bacterial suspen-sion was injected in each ball with a sterile disposable syringe (1 ml) (Beijing Fornurse Medical Equipment Co., Ltd. Ltd, China).

The infecting dosage was 105 CFU of S. aureus 209P per animal. The implant was inserted through a layer-by-layer dissection of soft tissues down to the required depth. Further on, the wound was sutured in layers for the above-mentioned period. The im-plant was removed surgically, and the tested material was inserted into the resulting cavity. In the control group, the animals did not have the material sample injected into the wound. The tissue defect above the experimental cavity was layer-by-layer sutured with apposition interrupted sutures. The surface area (S) and volume (V) of the cavity were calculated using the following formulas: S=4πr2, V=4/3πr3, where r is the radius of the implant, π = 3.14.

Throughout the study, the skin sutures were treated with an antiseptic (the trade name of the antiseptic is not mentioned here thus aiming to avoid a conflict of interest).

Experimental samples of multifunctional chitosan-based wound coatings were developed and synthesized on the basis of the National Research

S U R G E R Y


Center “Kurchatov Institute” (Table 1). The specific issue about the materials was the density and the pore orientation — isotropic or vertical.

Table 1. Technological features of chitosan samples used through the experiment

Major study outcomes and registration methodsThe main outcome of the study was the identifi-

cation of the most appropriate sample in terms of its

SamplePorosity Wall thickness

range Pore size range Elasticity modulus Deformation in compression

Offset yield strength

[%] [nm] [µM] [MPa] [%] [kPa]#1 98 350-1000 20-45 0.749 44.32 29.70#2 98 600-1200 70-200 0.369 50.14 24.15

Table 1 offers a view at the major parameters featured by the samples under study.

All experimental surgical interventions per-formed on laboratory animals were done in aseptic conditions and under general anesthesia. A combina-tion of drugs (atropine sulfate, prednisone, Sedami-dine, Telazol) was used as the anesthetic.

To reduce the general anesthesia time, upon com-pletion of the respective manipulations, the animals were injected with Antimedin. To relieve the pain syn-drome within the postoperative period (Day 1), the ani-mals were injected with Flexoprofen (intramuscular).

Inclusion criteriaFor the purposes of the study (through all of its

stages), male rats, as well as male rabbits were used with no external signs of diseases and anatomical disorders, that were quarantined in the retainer unit of the academic & production department of the Kuban State Medical University. The study involved animals that developed purulent wounds with standard signs of purulent inflammation during the simulation period.

Environment of studyThrough the entire study, including postoperative

periods, the animals were monitored with free access to water and food, in accordance with Russian State Regulatory Standards 33044-2014 “Principles of Good Laboratory Practice”. The study was carried out on the premises of the training and production department of the Kuban State Medical University (Krasnodar, Russia).

Duration of studyEach stage of the study went on for 28 days, tak-

ing into account the 7 days required for the wound cavity development. Measurements, diagnostics, and sampling of the material for histomorphological examination at all the stages were done on Days 7, 14, and 21.

parameters in the experimental groups, as well as the observed differences in the wound process dynamics between the control group and the experimental one.

An additional expected outcome was compli-ance with the data obtained through non-invasive ultrasound imaging of the wound defect area, in order to study changes in the structure of the tested mate-rial samples undergoing biodegradation, as well as the dynamics of changes affecting tissues in the area of the experimental wound.

The study involved analyzing the wound process parameters (reduction of the wound volume, heal-ing rate, sample biodegradation rate, complications, histomorphological composition of the wound) while using various samples of the designed wound coat-ing. Visual assessment of the wound process course included keeping track of the timing of reduction of edema, hyperemia of the tissues around the wounds, as well as any discharge from the wound.

The tissues were subjected to histomorpho-logical assessment following the generally accepted algorithm. The explanted tissues were fixed for 3–5 days in a 10 % neutral formalin solution (Histolab, Sweden), washed in running water for 60 minutes. The materials were processed subject to the stand-ard procedure employing an automatic method on a Leica TP1020 histoprocessor (Germany). The production of paraffin blocks with samples of the studied materials was carried out using a modular installation Leica EG1150H (Germany). A rotary microtome Leica RM2235 (Germany) was used for slicing the preparations, while the obtained sections (thickness — 5 microns) were stained with hema-toxylin and eosin according to the standard method. The micro-preparations were studied on an Olym-pus XH41 microscope ( Japan). The morphological evaluation of the created materials structure was done in the Versa 3D SEM/FIB DualBeam scanning election microscope (FEI, USA) with various ac-celerating voltages, equipped with a Gaseous Second-

S U R G E R Y


ary Electron Detector, a Peltier-cooled table and an option of using the ambient mode.

Ultrasound examination of the tissues was carried out with a Mindray M7 ultrasound scanner, using a high-frequency linear ultrasound sensor L 12-4s (China), with an operating frequency of 6–10 MHz — on the day the surgery was performed and then two days later, until the animal was removed from the experiment, in the following modes: in the color Dop-pler mapping mode using a pulse-wave Doppler as well as in the constant-wave Doppler mode.

Statistical analysisAt Stage 1, 2 samples of wound covering were ex-

amined in different modifications, for which 4 experi-mental and 1 control groups were formed, 45 animals (conventional male rats) in each group. Within the control term time, 15 animals within each group were removed from the experiment.

Stage 2 involved studying 2 types of wound coating samples of (# 1 and # 2) based on the criteria of porosity, mechanical strength and biodegradation terms. The properties of the samples were studied in 2 experimental and one control groups, each including 45 animals (linear male Wistar rats). Within the con-trol period, 15 animals of each group were removed from the experiment.

At the final stage, male rabbits were used, which were broken into 2 experimental and 1 control groups, 9 animals in each. In each control period, similar to the previous ones, 3 animals of each group were removed from the experiment.

The statistical processing of the study results was done on a personal computer in the Windows 10 op-erating system using the STATISTICA 6.1 (StatSoft, Inc., USA) and Excel (Microsoft Office 2010) soft-ware. The hypothesis of the cumulative distribution normality in the samples was tested relying on the Shapiro-Wilk and Kolmogorov-Smirnov criteria. The statistical significance level was set at p<0.05. The differences between the quantitative parameters with a normal distribution were evaluated through the Stu-dent’s t-test, whereas independent samples were evalu-ated relying on the nonparametric Mann-Whitney test. The differences in all cases were considered statistically significant at p<0.05. The significance level of the relationship between the two qualitative variables was tested through Pearson’s Chi-square (χ2) test.

R E S U L T SMajor results of study

Preliminary screening studies carried out at Stage 1 of this experiment revealed 2 most promising types of wound coatings made of modified chitosan, which

featured fundamental differences, both physical and chemical. This allows differential change in the tactics of purulent wound local treatment, depending on the specific task.

Stage 2 of the experiment implied studying the samples of wound coatings selected through Stage 1 following the criteria of porosity, mechanical strength and biodegradation time. The properties of the sam-ples were studied in conditions of a purulent wound. The result of the study revealed certain advantages demonstrated by sample # 1. Its biodegradation proc-ess was uniform; the distribution in the wound cavity was equal, which contributed to a smooth therapeutic effect in all parts of the wound cavity.

The assessment of the biodegradation degree of the samples studied at Stage 2 in % ratio to the initial volume revealed a statistically significant difference in biodegradation between samples # 1 and # 2. Sample # 2 underwent biodegradation 7 times as fast (p<0.01) and by Day 9 was basically undetectable within the operation area (Fig. 1).

Fig. 1. Residual wound coverage volume (% of original) in the target time frame – p<0,01.

The histological representation, following the introduction of a test material sample, is a developed abscess in the muscle tissue, where the walls consist of 2 layers — a pyogenic membrane (up to 0.3 mm) and a developing connective tissue capsule (up to 1.5 mm). The abscess cavity contained a partially biode-graded sample distributed evenly. The muscle tissues surrounding the capsule featured an edema with an increased number of full-blood capillaries, with certain signs of stasis and lymph-macrophage infiltration (Fig. 2).

Stage 3 was carried out in order to study the chitosan-based multifunctional wound coating prop-erties in larger purulent wounds affecting soft tissues.

S U R G E R Y


The area chosen to develop the wound cavity was the shoulder-scapular region, whereas the trapezius muscle tissue was chosen as the depth of the simulated cavity.

The polyvalent potential of using the developed samples is due to the experimentally proven proper-ties featured by the selected samples: proper frame function (sufficient density), due bioadhesive capacity, biodegradability and biocompatibility.

All these properties identified in experimental samples were to be found in them in different propor-tions, which could be accounted for by the specifics of their production technology — the share chitosan in the base solution being the first one, as well as the molecular weight of the sample; features of the pore

a) Appearance of sample No. 3, with a magnification of x4

b) Fragments of sample no. 3; homogeneous biodegradation (7 days)

Fig. 2. a) — Appearance and histomorphological picture of sample No. 3, with signs of homogeneous biodegradation on the 7th day after its introduction into the experimental wound b) — (1 — homogeneous biodegradation of the material; 2-pyogenic capsule).

internal orientation and the ratio of the size and thick-ness of the walls separating them.

Given that, a programmable change in the initial parameters’ ratio of the raw stuff and the use of various options for manufacturing chitosan samples, allow al-tering the finally obtained properties of the developed wound coatings as listed above.

Extra results of studyAt Stage 3, the experiment was carried out rely-

ing on intravital ultrasound of the wound defect area (Fig. 3). The purpose of the ultrasound study was to investigate possible visualization of changes affecting the structure of the test material samples undergo-ing biodegradation, as well as the dynamics of tissue changes at the experimental wound.

The use of ultrasound to monitor the experimen-tal wounds through the experiment, done in order to ensure a more detailed study of the wound cavity and the surrounding tissues status could be explained by a number of reasons. At the stage of developing the wound cavity, the ultrasound method allowed visual-izing the implant inserted into the soft tissues as well as the blood flow in the surrounding tissues, which helped assess the location of the implant in relation to the surrounding anatomical formations, and was of great importance for standardizing the process of the wound cavity development. In view of the dynamics, the ultrasound study allowed identifying the degree of the wound cavity capsule development, any wound discharge inside the cavity, as well as the blood flow status at the surgical intervention area.

At the further stages of the experiment, the meth-od of ultrasound examination of the experimental wound area containing a coating sample allowed not only visualizing biodegradation of the tested material, yet also identifying the major ultrasound semiotics of soft tissue damage – inflammatory infiltrate at differ-ent stages of its progress, muscle fiber imbibition, the appearance of a purulent substrate and signs pointing at the beginning reparation.

The verification of the data obtained through the ultrasound examination was done by studying and comparing the biopsy material obtained when removing the animals from the experiment within the above-described control periods. This method helped us arrive at non-invasive assessment of the wound proc-ess status when using the selected samples of wound coatings, as well as it helped visualize the samples of the studied material, which were undergoing biodegra-dation (Fig. 4).

Following the introduction of chitosan samples into the muscle layer, they were visualized as forma-tions featuring clear and even contours and a homoge-

S U R G E R Y


and echogenicity visualized. Vascular structures of muscle tissue were visualized as single blood flow loci revealing a spectrum of arterial and venous blood.

By Day 7 into the observation, the size of the wound coating samples regressed. What is more, sample # 2 had nearly completely biodegraded: the size of the wound cavity shrank down to 5–10% of its initial size. In the remaining wound cavity, a small amount of exudate (up to 0.25 ml) was to be observed. During that, the size of sample # 1, which was the denser one, as well as the size of the cavity it was placed in, shrank down to 30–35% of its initial size. The peripheral area of the remaining wound cavity featured denser hyperechogenic and homogeneous layers of muscle fibers of various thicknesses (2.5 mm to 12 mm) appearing there.

There was also an increase observed in the blood flow speed within the arterial and venous network – Vmax art — 25–30 cm/s; Vmax veins – 15–17 cm/s.

a) Conducting an ultrasound scan at the stage of introducing a polymer implant into soft tissues at stage 3 of the experiment

b) Assessment of the blood supply to the surrounding tissues (1) (2)

Fig. 3. a) Ultrasound at the stage of introducing a polymer implant into soft tissues. b) — Assessment of blood supply of tissues (2) surrounding implant (1)

neous structure with the effect of distal pseudo-rein-forcement, the size being up to 20–20–20 mm.

Along the periphery of the implanted sample, there were vital tissues with clear ultrasound structure

a) Introduction of a sample of the test material (1) into the formed experimental wound

b) Ultrasonic location of the sample (1) of the test material in soft tissues

Fig. 4. Comparison of а) visual and b) CD images of tissues and samples of the drug, when performing various stages of the study

S U R G E R Y


The study conducted on Day 14 into the experi-ment revealed that infiltrative changes affecting the muscle tissue surrounding the experimental wound cavity featured different stages of inflammation. There were signs of dense and loose infiltrate (layer thickness ranging from 1.7 mm to 9.8 mm) visualized, as well as the muscle fiber imbibition, with an increase in its volume (from 4.4 mm to 12.8 mm). The cavity of the wound defect, where the denser sample # 1 was imple-mented, decreased to 1.2 x 4.3 x 1.7 mm (H x W x D), with some exudate identified, which manifested itself as anechogenic strips on the periphery of the sample residues (up to 0.5 ml).

Upon introducing sample # 2, the residual effects in the experimental wound were visualized as loose-fibrous (hyperechogenic) formations with a diameter of up to 2.5 mm, which featured signs of infiltrative changes in the surrounding tissues. The maximum speed of the venous blood flow in the infiltrate zone was Vmax veins — 5 cm/s, which almost matched the initial value.

There was also a clear visualization of the bounda-ries separating the muscle-aponeurotic layers in the respective area, with a differentiation of a striated muscle tissue pattern.

To further study the tested wound coating sample biodegradation, we relied on the scanning electron microscopy method. On Day 7 into the experiment, samples # 1 were obtained. The result of the study con-firmed the theoretical calculation of the step-by-step sample biodegradation with the capillary (sorption) properties preserved (Fig. 5).

Adverse eventsNo adverse events were observed through the

experimental study.

D I S C U S S I O NThe developed wound coatings featured proper

framework functions, as well as due bioadhesive capac-ity, biodegradability and biocompatibility.

The study results obtained through investigating the properties of the developed chitosan-based wound coatings correlate with the literature data in terms of comparing properties like bioadhesive capacity and biodegradability [21]. The experimental data appear comparable with the results other researchers obtained — there was confirmation obtained for the calculated qualities of the impact that the respective samples had on the surrounding tissues, in view of the specifics of the wound defect occurrence and specific wound heal-ing processes, as well as there was potential identified for ensuring a proper microclimate: controlled pH, humidity; thermal insulation of the wound; creating a

barrier against microorganisms; ensuring due sorption activity of the material; atraumatic capacity in terms of granulations [15–17].

A comparison of the developed wound coatings revealed that various synthesis technologies allowed controlling their physical and chemical properties as well as possible use of these coatings within various treatment schemes not only when dealing with issues of infectious origin, yet also when treating other soft tissue lesions.

C O N C L U S I O NThe chitosan-based wound coatings studied

through a multi-stage experiment offered proof to the properties predicted while developing the materials, namely, a stable adhesive capacity to biological tissues and the framework function.

b) Internal — with preserved structure (1) and external — partially biode-graded (2) surfaces of the test material in soft tissues

Fig. 5. Investigation of the biodegradation process by scanning electron microscopy

a) Cellular structure preserved in the sample

S U R G E R Y


Sample # 1 featured moderate strength, elasticity, as well as a denser structure and, respectively, a lower level of biodegradation (bioresorption on Day 14), which determined its capacity to maintain the desired shape and the initial dimensions for a long time. This, in turn, allowed the sample to effectively perform the framework functions and ensure proper drainage of the pathology focus. The mentioned features revealed that the sample could offer a promising choice when dealing with purulent wounds.

Sample # 2 was of lower strength, elasticity, density, as well as a higher adhesion degree (compared to sample # 1), with complete biodegradation achieved on Day 7. In view of the test sample features, it can prove of the best use in cases with no purulent inflam-mation and with granulating wounds. Besides, sample # 2 revealed a capacity to transform into a gel with a highly ordered internal structure of the micellar type, which allowed introducing another function – the use as a carrier matrix for the drugs introduced into its structure. This condition will allow expanding the indications for use.

The parallel ultrasound examination technique employed to study experimental wound areas pro-duced an expected positive effect. The match between the data obtained during non-invasive visualization of the analyzed samples and the surrounding tissues, with the data obtained through histomorphological studies allowed reducing both the number of invasive proce-dures and the number of animals to be involved in the study process.

E T H I C S P R I N C I P L E S C O M P L I A N C EThe study described above was granted approval

at the meeting of the Independent Ethics Committee, the Kuban State Medical University (Protocol # 63 of May 21, 2018) and was carried out within the period of 2019-2021.

F U N D I N GThe authors herewith claim no sponsorship used

for the study.

R E F E R E N C E S1. Belik B. M., Chumburidze I. P., Shtilman M.

Yu., Yavruyan O. A, Savchenko Yu. P. Complex treatment of patients with purulent-necrotic complica-tions in neuropathic form of diabetic foot syndrome. Surgery. Journal them. N.I. Pirogov. 2020; (4): 81–87. https://doi.org/10.17116/hirurgia202004181

2. Sepsis: classification, clinical and diagnostic concept and treatment. / Ed. Gelfand B.R. – Moscow: MIA; 2017. (In Russ).

3. Chiang N, Rodda OA, Sleigh J, Vasudevan T. Effects of topical negative pressure therapy on tissue

oxygenation and wound healing in vascular foot wounds. Journal of Vascular Surgery. 2017 Aug 1; 66(2): 564–571. DOI: 10.1016/j.jvs.2017.02.050

4. Belov A.A., Vanyushenkova A.A., Dosadina E.E., Khanafina A.A. New textile dressings based on biodegradable polymers containing proteinases for the treatment of wounds and burns. Wounds and wound infections. 2018; 5 (1): 16–26. DOI: 10.25199/2408-9613-2018-5-1-16-26

5. Yan D, Liu S, Zhao X, Bian H, Yao X, Xing J, et al. Recombinant human granulocyte macrophage colony stimulating factor in deep second-degree burn wound healing. Medicine (Baltimore). 2017; 96(22): 6881. DOI: 10.1097/md.0000000000006881

6. Katorkin S.E., Bystrov S.A., Lisin O.E., Rozanova A.A., Bezborodov A.I. Evaluation of the effectiveness of the use of modern dressings in the complex treatment of purulent wounds. Inpatient Replacement Technologies: Outpatient Surgery. 2019; 1–2: 146–152. DOI: 10.21518/1995-1477-2019-1-2-146-152

7. Mueller T.C. Intra-operative wound irrigation to reduce surgical site infections after abdominal surgery: a systematic review and meta-analysis. Langenbecks Arch. Surg. 2015; 400 (2): 167–181. DOI: 10.1007/s00423-015-1279-x

8. Huang W., Wang Y., Huang Z., Wang X., Chen L., Zhang Y., Zhang L. On-Demand Dissolvable Self-Healing Hydrogel Based on Carboxymethyl Chitosan and Cellulose Nanocrystal for Deep Partial Thickness Burn Wound Healing. ACS Appl Mater In-terfaces. 2018;10 (48): 41076–41088. DOI: 10.1021/acsami.8b14526

9. Proud D., Holden D., Capstick R., et al. Are we getting necrotizing soft tissue infections right? A 10-year review. ANZ J Surg. 2014; 84; 6; 468–472. DOI: 10.1111 / ans.12412

10. Atkin L. Understanding methods of wound debri-dement. Br J Nurs. 2014 Jun 26-Jul 9;23(12): S10–2, S14–5. doi: 10.12968/bjon.2014.23.sup12.S10

11. Katorkin S.E., Bystrov S.A., Lisin O.E., Ro-zanova A.A., Bezborodov A.I. Evaluation of the effectiveness of the use of modern dressing materi-als in the complex treatment of purulent wounds // Inpatient replacement technologies: Outpatient surgery. 2019. №1–2:146–152. DOI: https://doi.org/10.21518/1995-1477-2019-1-2-146-152

12. Fakayode OJ, Tsolekile N, Songca SP, Oluwa-femi OS. Applications of functionalized nanomateri-als in photodynamic therapy. Biophys. 2018; 10: 1: 49–67. DOI: 10.1007/s12551-017-0383-2

13. Childs DR, Murthy AS. Overview of Wound Healing and Management. Surg Clin North Am. 2017;97(1):189–207. DOI: 10.1016/j.suc.2016.08.013

14. A. A. Belov, A. A. Vanyushenkova, E. E. Dosadina, A. A. Khanafina. New textile dressings based on biodegradable polymers containing protein-ases for the treatment of wounds and burns. Wounds and wound infections. Journal them. prof. B. M. Ko-

S U R G E R Y


styuchenka. 2018; 5 (1): 16–26 DOI: 10.25199/2408-9613-2018-5-1-16–26

15. Mohammadtaghi Vakili, Mohd Rafatullah, Babak Salamatinia, Ahmad Zuhairi Abdul-lah, Mahamad Hakimi Ibrahim, Kok Bing Tan, Zahra Gholami, Parisa Amouzgar, Ap-plication of chitosan and its derivatives as adsorbents for dye removal from water and wastewater: A review, Carbohydrate Polymers,Volume 113,2014,Pages 115–130,ISSN 0144-8617, https://doi.org/10.1016/j.carbpol.2014.07.007

16. Ahmed S, Ikram S. Chitosan based scaffolds and their applications in wound healing. Achievements in the Life Sciences. 2016;10(1):27–37. https://doi.org/10.1016/ j.als.2016.04.001

17. Mohandas A, Deepthi S, Biswas R, Jayakumar R. Chitosan based metallic nanocomposite scaffolds as antimicrobial wound dressings. Bioact Mater. 2018;3(3):267–277. https://doi.org/10.1016/j.bioact-mat. 2017.11.003

18. Mayorova A.V., Sysuev B.B., Khanalieva I.A., Vikhrova I.V. Modern assortment, properties and prospects for improving dressings for the treatment of wounds. Pharmacy and pharmacology. 2018;6(1): 4–32. DOI: 10.19163/2307-9266-2018-6-1-4-32

19. Julia Radwan-Pragłowska, Marek Piątkowski, Volodymyr Deineka, Łukasz Janus,, Viktoriia Korniienko, Evgenia Husak, Viktoria Holubnycha, Iryna Liubchak, Vyacheslav Zhurba, Aleksandra Sierakows-ka, Maksym Pogorielov, Dariusz Bogdał. Chitosan-Based Bioactive Hemostatic Agents with Antibacterial Properties-Synthesis and Characteriza-tion. Molecules 2019, 24(14), 2629; DOI: 10.3390/molecules24142629

20. Singh R., Shitiz K., Singh A. Chitin and chitosan: biopolymers for wound management. Int. Wound. J. 2017; 14(6): 1276–1289. DOI:10.1111/iwj.12797

21. Oltarzhevskaya N.D., Korovina M.A., Krichevsky G.E., Shchedrina M.A., Egorova E.A. Possibilities of using polysaccharides in the treatment of wounds. Wounds and wound infec-tions. Journal named after prof. B.M. Kostuchenka. 2019;6(2):24–31. https:// DOI: 10.25199/2408-9613-2019-6-2-24-31

22. Yi-Wen Wang, Chuan-Chieh Liu, Juin-Hong Cherng, Chien-Seng Lin, Shu-Jen Chang, Zhi-Jie Hong, Cheng-Che Liu, Yaw-Kwan Chiu, Sheng-Der Hsu, Hung Chang/ Biological Effects of Chitosan-Based Dressing on Hemostasis Mecha-nism. Polymers 2019, 11(11), 1906; DOI: 10.3390/polym11111906

23. Zhang Hu, Sitong Lu, Yu Cheng, Songzhi Kong, Sidong Li, Chengpeng Li andLei Yang/ Investigation of the Effects of Molecular Parameters on the Hemostatic Properties of Chitosan. Molecules 2018, 23(12), 3147; https://doi.org/10.3390/mole-cules23123147 Faculty of Chemistry and Environmen-tal Science, Guangdong Ocean University, Zhanjiang

524088, China; Authors to whom correspondence should be addressed.

24. Gumenyuk A., Ushmarov D.I., Gumenyuk S.E. Potential use of chitozan-based multilayer wound covering in dental practice. Archiv EuroMedica. 2019. Vol. 9. № 3. Р. 76–80. https://doi.org/10.35630/2199-885X/2019/9/3.24

25. Karpyuk V.B., Perova M.D., Gilevich I.V., Sevostyanov I.A. Cell-potentiated regenerative technologies for restoring jaw bone tissues in case of odontogenic inflammatory & destructive process // Archiv EuroMedica. 2019. Vol. 9. № 2. P. 140–146. https://doi.org/10.35630/2199-885X/2019/9/2/140

26. Karpyuk V.B., Perova M.D. Innovation-based ap-proach in reconstruction of reduced jaw alveolar ridge bone using cell regeneration technologies // Archiv EuroMedica. 2019. Vol. 9. № 2. P. 147–155. https://doi.org/10.35630/2199-885X/2019/9/2/147

27. Bairikov I.M., Gaivoronskaya T.V., Dedikov D., Stolyarenko P.Yu. Reconstruction of man-dibular defects using individual vascularized autografts combined with macroporous titanium fiber material // Archiv EuroMedica. 2021. Vol. 11. № 1. P. 147–159. https://doi.org/10.35630/2199-885X/2021/11/1.32

S U R G E R Y


PREVENTION OF POST-ERCP PANCREATITIS IN HIGH-RISK PATIENTS

A B S T R A C T — Aim. To study the efficacy of thoracic epidural analgesia (TEA) for the prevention of post-ERCP pancreatitis in high-risk patients.Materials and methods. A parallel, blinded, randomized study. The first (TEA group) group included patients (n = 98) in whom thoracic epidural analgesia was used during endoscopic transpapillary interventions (ETI), the second (OAI group) group included patients (n = 97) in whom opioid analgesics and indomethacin (per rectum).Results. The study showed that acute pancreatitis was diagnosed significantly less frequently in patients with the TEA group than in patients with the OAI group (p = 0.0135). If in the TEA group post-ERCP pancreatitis (PEP) was verified in 3.1% (3/98) patients, in the OAI group — in 12.4% (12/97) patients.When TEA was used in high risk patients of developing post-ERCP pancreatitis, its incidence decreased from 23.3% (10/43) to 4.4% (2/46) observations (p = 0.0095).Conclusion. The use of TEA is an effective and justified method of prevention in patients at high risk of developing post-ERCP pancreatitis. In patients with a low risk of developing this complication, the use of TEA is inappropriate due to the invasiveness of the method.

K E Y W O R D S — therapeutic ERCP (endoscopic retrograde cholangiopancreatography), prevention of post-ERCP pancreatitis, thoracic epidural analgesia.

Mikhail Turovets , Alexander Popov ,Viktor Mandrikov , Yuriy Vedenin ,Andrej Ekstrem

Volgograd State Medical University, Volgograd, Russia

[email protected]

Received 27.03.2021 Received in revised form 29 April 2021; Accepted 3 Mai 2021

I N T R O D U C T I O NThe use of endoscopic transpapillary interven-

tions (ETI) to correct biliary hypertension syndrome has significantly improved the results of treatment in patients with pathology of the duodenopancreatobil-iary zone.

At the same time, ETI are complex interven-tions with unpredictable consequences. During the perioperative period, severe and sometimes fatal complications may occur, such as post-ERCP pancrea-titis (PEP), bleeding from the papillotomy area, and retroduodenal perforation [1, 2, 3, 4].

According to many researchers, the incidence of post-ERCP pancreatitis is within 1–40% of observa-

tions and depends on many factors: the nature of the disease, the type of endoscopic intervention, the patient's age [2]. Such factors as young age, sphincter of Oddi dysfunction, the use of balloon dilatation for ETI, the absence of jaundice increase the risk of its development [3, 5, 6, 10]. To date, mortality in post-ERCP pancreatitis reaches 3–10%, and with develop-ing infected pancreatic necrosis can reach 25–80% of cases [1, 2, 7, 8, 11].

In recent years, there have been positive responses from the research community on the effectiveness of non-steroidal anti-inflammatory drugs (diclofenac and indomethacin) and thoracic epidural analgesia for the prevention of PEP in ETI [1, 12]. Thus, we decided to conduct a comparative analysis to test the effective-ness of these two methods of preventing post-ERCP pancreatitis in high-risk patients.

M A T E R I A L S A N D M E T H O D S Our parallel, unblinded, randomized study was

approved by Volgograd Regional Ethics Committee. All participants signed an informed consent.

223 ETI were conducted from January 2019 to December 2020. All patients were hospitalized. In 13 (4.9%) patients, endoscopic intervention was per-formed for diagnostic purposes (excluded from the study). In 9 out of 223 (4.0%) patients, signs of acute pancreatitis were diagnosed before the intervention (excluded from the study). One patient refused to participate in the study. A total of 200 patients were included in the study.

Prior to the formation of the database, the study design, inclusion and exclusion criteria were determined.

Inclusion criteria:1. Completed therapeutic ETI;2. Prior to ETI, the patient had no clinical signs

of acute pancreatitis.Exclusion criterion: a complication (retroduo-

denal perforation, massive bleeding, detachment of the Dormia basket) was diagnosed during ETI and required an urgent surgical intervention.

All subjects were divided into two groups (100 patients each). The first (TEA group) group included patients in whom TEA was used during ETI, the patients of the second (OAI group) group received a narcotic analgesic and indomethacin.

Puncture and catheterization of the epidural space was performed according to the standard tech-


http://dx.doi.org/10.35630/2199-885X/2021/11/2/17

https://orcid.org/0000-0003-0793-5098

https://orcid.org/0000-0003-2241-8144

https://orcid.org/0000-0002-5890-8500

https://orcid.org/0000-0001-9004-7694

https://orcid.org/0000-0002-0453-6158

http://dx.doi.org/10.35630/2199-885X/2021/11/2/17


nique at the level of the VII–VIII thoracic vertebrae. 20 minutes before ETI, a solution of ropivacaine 0.5% – 10 ml was injected into the epidural space.

In the TEA group, two patients were excluded from the study due to unsuccessful attempts to insert a catheter into epidural space (1 case) and retroduodenal perforation (1 case). In the OAI-group, three patients were excluded from the study: due to massive bleeding (1 case), retroduodenal perforation (1 case), and reten-tion of the Dormia basket (1 case), which required surgical interventions.

As a result, the results of treatment of 195 patients (98 patients of the TEA group and 97 patients of the NAI group) were analyzed.

The studied indicators (demographic data, the nature of the disease, the results of laboratory or in-strumental research, outcomes, etc.) were entered into the database within 10 days after the intervention.

Statistical analysis of the data was performed using nonparametric tests (OR and Fisher's exact test). The groups were divided into subgroups taking into account age, sex, nature of the disease, severity of con-comitant pathology, types of ETI. For each of the sub-groups, the odds ratio (OR) was calculated with a 95% confidence interval (95% CI). A statistically significant difference between the study groups (subgroups) was considered p ≤ 0.05 (an indicator of statistically signifi-cant differences) or when the 95% confidence interval (CI) did not include 1. Statistical processing of the data was carried out using a set of statistical programs Statistica 10.0 (StatSoft Inc., USA).

R E S U L T S A N D D I S C U S S I O N The demographic data of the patients are pre-

sented in Table 1. In the TEA-group, the average age of the patients was 61.4 ± 1.3 years, in the OAI-group — 60.9 ± 1.2 years. There were no statistically signifi-cant differences in the number of women and patients of young age (under 60 years) and more advanced age (over 60 years) (OR 0.94 [95% CI, 0.53–1.66]) and (OR 1.06 [95% CI, 0.60–1.88 ], respectively).

The study groups did not differ in other indica-tors either. In the TEA group and the OAI group, there was a commensurate number of patients with tumors of the hepatopancreatobiliary zone (33.7% [33/98] versus 33.0% [32/97] cases, OR 1.03) and with jaundice (38.8% [38/98] versus 38.1 % [37/97] cases, OR 1.03); with choledocholithiasis (36.7% [36/98] versus 38.1% [37/97] cases, OR 0.94) and with sphincter of Oddi dysfunction (SOD) (12.2% [12/98] versus 11.3% [11/97] observations, OR 1.09).

Concomitant diseases were detected in 58.2% (57/98) of patients in the TEA group and in 61.9% (60/97) of patients in the OAI group (OR 0.85 [95%

CI, 0.48–1.53]). The study groups also did not differ in the number of patients with severe comorbidity (ASA IV) (OR 1.13 [95% CI, 0.41–3.08]). The major-ity of patients in this category were diagnosed with diseases of the cardiovascular system (84.2% [48/57] of patients with the TEA group and 85.0% [51/60] of patients with the OAI group).

The study groups also did not differ on other vari-ables. Based on the above, the study groups represent-ed patients with similar baseline parameters, and this could not affect the reliability of the results.

Depending on the nature of the disease, various types of endoscopic transpapillary interventions were used. The study groups did not differ in this indicator either. In the TEA group and the OAI group, endo-scopic papillosphincterotomy was equally often used (in 82.7% versus 81.4% of cases (OR 1.09 [95% CI, 0.52–2.27])) and balloon dilation (in 18.4% versus 20.6% of cases (OR 0.87 [ 95% CI, 0.42–1.77])), bil-iary-stone extraction (in 51% versus 53.6% cases (OR 0.90 [95% CI, 0.51–1.59])) and lithotripsy (in 9.2% versus 10.3% of cases (OR 0.88 [95% CI, 0.34–2.29])), installation of biliary stent (31.6% versus 28.9% of cases (OR 1.14 [95% CI, 0.39–3.32])) and installation of pancreatic stent (19.4% versus 22.7% of cases (OR 0.82 [95% CI, 0.41–1.65])).

During this study, patients in the study groups were diagnosed with 15 (7.7% [15/195]) cases of PEP development. In 80.0% (12/15) of observations, this complication was registered in patients of the OAI-group (Table 2). The incidence of PEP in the TEA group was 3.1% (3/98), and in the OAI group — 12.4% (12/97) of observations, which is a statistically signifi-cant difference between the study groups (p = 0.0135).

The vast majority of cases of PEP in patients of the TEA group and OAI group were mild (66.7% [2/3] and 58.3% [7/12] observations, respectively) and their clinical manifestations could be stopped within 3 days. One of three patients (33.3%) of the TEA group and 4 of 12 (33.3%) patients of the NAI-group needed to continue anti-pancreatic therapy for up to 7 days. Taking into account the clinical picture, the data of laboratory and instrumental studies, in the NAI-group, pancreatonecrosis was verified in 1 (1.0%) patient. Symptoms of pancreatic necrosis in this patient devel-oped rapidly and were characterized by total damage to the pancreas, complicated by the development of multiple organ failure, which led to death.

The incidence of post-ERCP pancreatitis in study subgroups different in gender, age, disease nature, se-verity of concomitant pathology and types of interven-tion is shown in Table 3.

A statistically significant decrease in the incidence of PEP with the use of TEA was found in patients



VariableTotal, n (%) OR

(95% CI)TEA group (N=98) OAI group (N=97) Age 18-40 y 9 (9.2) 8 (8.2) 1.13 (0.41-3.08) 41-60 y 38 (38.8) 40 (41.2) 0.90 (0.51-1.61) 61-80 y 39 (39.8) 36 (37.1) 1.12 (0.62-2.00) ˃80 y 12 (12.2) 13 (13.5) 0.90 (0.39-2.11)Gender Woman 59 (60.2) 61 (62.9) 0.89 (0.50-1.60)Context Jaundice 38 (38.8) 37 (38.1) 1.03 (0.57-1.84) Common bile duct stones 36 (36.7) 37 (38.1) 0.94 (0.52-1.69) SOD 12 (12.2) 11 (11.3) 1.09 (0.45-2.63) Common bile duct stones and SOD 14 (14.3) 15 (15.5) 0.91 (0.41-2.02)

Tumor 33 (33.7) 32 (33.0) 1.03 (0.56-1.88) Calculous pancreatitis 3 (3.1) 2 (2.1) 1.50 (0.24-9.37)ASA grade IV 9 (9.2) 8 (8.2) 1.13 (0.41-3.08) III 21 (21.4) 22 (22.7) 0.93 (0.47-1.84) I and II 68 (69.4) 67 (69.1) 1.01 (0.55-1.88)Procedural Biliary sphincterotomy 81 (82.7) 79 (81.4) 1.09 (0.52-2.27) Balloon dilation 18 (18.4) 20 (20.6) 0.87 (0.42-1.77) Biliary-stone extraction 50 (51.0) 52 (53.6) 0.90 (0.51-1.59) Installation of biliary stent 31 (31.6) 28 (28.9) 1.14 (0.61-2.12) Lithotripsy 9 (9.2) 10 (10.3) 0.88 (0.34-2.29) Installation of pancreatic stent 19 (19.4) 22 (22.7) 0.82 (0.41-1.65) Naso-biliary drainage 8 (8.2) 7 (7.2) 1.14 (0.39-3.32)

Table 1. Selected subject and procedural characteristics of patients

Note. *P < 0.05, statistically significant; ASA — American Society of Anaesthesiologists; SOD — sphincter of Oddi dysfunction

Table 2. The incidence of post-ERCP pancreatitis in patients of the study groups

Post-ERCP pancreatitis TEA group (N=98)

OAI group (N=97) P, Fisher

Mild, n(%) 2 (2.1) 7 (7.2) 0.0823Moderate, n(%) 1 (1.0) 4 (4.2) 0.1811Severe, n(%) 0 (0.0) 1 (1.0) 0.4974Total, n(%) 3 (3.1) 12 (12.4) 0.0135*

under 60 years of age (OR 0.18 [95% CI, 0.04–0.93]) and with the use of endoscopic papillosphincterotomy (OR 0.17 [95% CI, 0.04–0.85]).

A significant (but statistically insignificant) decrease in the incidence of PEP with the use of TEA was found in patients of all study subgroups: in women

(OR 0.31 [95% CI, 0.08–1.24]), in patients with jaundice (OR 0.22 [95% CI, 0.02–2.20]) and without it (OR 0.22 [95% CI, 0.04–1.14]), in patients with choledocholithiasis and SOD (OR 0.21 [95% CI, 0.02–2.29]) and in patients with isolated choledocho-lithiasis (OR 0.24 [ 95% CI, 0.02–2.32]), after balloon



Table 3. The incidence of post-ERCP pancreatitis in patients of the study subgroups

Variable Post-ERCP pancreatitis n (%) OR

(95% CI)TEA group (N=98) OAI group (N=97) Age 18-40 y 1/9 (11.1) 2/8 (25.0) 0.38 (0.03-5.46) 41-60 y 1/38 (2.6) 6/40 (15.0) 0.15 (0.02-1.40) 61-80 y 1/39 (2.6) 3/36 (8.3) 0.29 (0.03-3.06) ˃80 y 0/12 (0.0) 1/13 (7.7) ---Gender Woman 3/59 (5.1) 9/61 (14.8) 0.31 (0.08-1.24) Men 0/39 (0.0) 3/36 (8.3) ---Context Jaundice 1/38 (2.6) 4/37 (10.8) 0.22 (0.02-2.20) Common bile duct stones 1/36 (2.8) 4/37 (10.8) 0.24 (0.02-2.32) SOD 0/12 (0.0) 1/11 (9.1) --- Common bile duct stones and SOD 1/14 (7.1) 4/15 (26.7) 0.21 (0.02-2.29)

Tumor 1/33 (3.0) 3/32 (9.4) 0.30 (0.03-3.22) Calculous pancreatitis 0/3 (0.0) 0/2 (0.0) ---ASA grade IV 0/9 (0.0) 1/8 (12.5) --- III 1/21 (4.8) 3/22 (13.6) 0.30 (0.03-3.31) I and II 2/68 (2.9) 8/67 (11.9) 0.22 (0.04-1.13)Procedural Biliary sphincterotomy 2/81 (2.5) 10/79 (12.7) 0.17 (0.04-0.85)* Balloon dilation 1/18 (5.6) 5/20 (25.0) 0.18 (0.02-1.77) Biliary-stone extraction 2/50 (4.0) 8/52 (15.4) 0.23 (0.04-1.18) Installation of biliary stent 1/31 (3.2) 4/28 (14.3) 0.20 (0.02-2.00) Lithotripsy 1/9 (11.1) 3/10 (30.0) 0.29 (0.02-3.67) Installation of pancreatic stent 0/19 (0.0) 2/22 (9.1) --- Naso-biliary drainage 1/8 (12.5) 2/7 (28.6) 0.36 (0.02-5.40)Total, n(%) 3/98 (3.1) 12/97 (12.4) 0.22 (0.06-0.83)*

Post-ERCP pancreatitisHigh risk patients

P, Fisher TEA group (N=46) OAI group (N=43)

Mild, n(%) 1 (2.2) 6 (14.0) 0.0454*Moderate, n(%) 1 (2.2) 3 (7.0) 0.2830Severe, n(%) 0 (0.0) 1 (2.3) 0.4831Total, n(%) 2 (4.4) 10 (23.3) 0.0095*

Table 4. Frequency of post-ERCP pancreatitis in high-risk patients

dilatation (OR 0.18 [95% CI, 0.02–1.77]), biliary-stone extraction (OR 0.23 [95% CI, 0.04–1.18]) and naso-biliary drainage (OR 0.36 [95% CI, 0.02–5.40]), etc.

In some subgroups, patients from the TEA group did not have a single case of post-ERCP pancreatitis,

namely, in patients over 80 years of age, in men, in patients with SOD, with calculous pancreatitis, with severe concomitant pathology (ASA IV), after installa-tion of pancreatic stent. There were no lethal outcomes in patients of the TEA group.



More often, the early postoperative period was accompanied by the development of AKI in women (in 10.0% [12/120] cases), in young (up to 60 years old) patients (in 10.5% [10/95] cases), and in patients with choledocholithiasis and SOD (in 17.2% [5/29] observations). According to our data, these factors increased the risk of post-ERCP pancreatitis, and the subjects, who combined two or more risk factors, were attributed to patients with a high risk of PEP [1]. In the TEA group, 46.9% (46/98) were identified, and in the OAI group — 44.3% (43/97) of patients in this category. Table 4 presents data on the incidence of post-ERCP pancreatitis in patients at high risk of developing this complication.

As shown in the table, when TEA was used in patients with a high risk of PEP, its incidence decreased from 23.3% (10/43) to 4.4% (2/46) observations (p = 0.0095).

In patients with a low risk of developing this complication, the use of both TEA and indometh-acin suppositories, as methods of preventing PEP, has shown commensurate efficacy. There was no statistical-ly significant difference in these study subgroups (3.7% (2/54) observations in the NAI subgroup versus 1.9% (1/52) cases in the TEA subgroup (p = 0.5143)).

Throughout the entire period of the use of thera-peutic ETI we havecontinued the search for methods of preventing the development of PEP. A lot of studies were carried out to study the preventive effect of the use of various groups of medicines (nitrates, san-dostatin, heparins, etc.) [4, 6, 7, 11]. For this purpose, various endoscopic tactics of endoscopic interven-tions were proposed for use [1, 9, 10]. But despite this, development of post-ERCP pancreatitis has remained the main problem of therapeutic ETI.

As a result of our study, reliable data were ob-tained on the effectiveness of of TEA for preventing the development of PEP in ETI. A decrease in the incidence of post-ERCP pancreatitis from 12.4% to 3.1% was shown. In the TEA group, PEP was mild to moderate in all cases. The use of TEA helps prevent the development of pancreonecrosis and poor out-comes in patients, which significantly increases the safety of ETI in patients with a high risk of PEP and severe comorbidity. An important fact is that the use of TEA, as noted by endoscopists, creates more com-fortable conditions for their work.

All patients were inpatient until the end of the study, so there was no data loss. The indicators were entered into the database in real time.

Of course, this method of prevention has its drawbacks. We refer to the main disadvantages: the need to involve a doctor who knows the methodology for conducting TEA; the invasiveness of the method

(despite its safety); restriction in use in patients with coagulopathies (for example, in patients with hepatic insufficiency); the possibility of using only in hospital-ized patients.

Of course, not all patients with therapeutic ETI need to use TEA. Obviously, in patients with a low risk of developing PEP, it seems sufficient to use a less invasive technique — indomethacin suppositories.

C O N C L U S I O N— The use of TEA is an effective and justified

method of prevention in patients at high risk of devel-oping PEP;

— In patients with a low risk of PEP, the use of TEA is inappropriate due to the invasiveness of the method.

In conclusion, we would like to note that this study does not question the effectiveness of other pre-vention methods recommended by different authors, but only complements them. Anyway, an individual approach to each patient is required.

Conflict of interestsThe authors state that they have no conflict of

interests.

ContributorsMIT and VVM collected, analysed, and in-

terpreted data and made the figures. ASP did the literature review and collected data. AVE and YuIV collected data and made the figures. MIT and VVM interpreted and analyzed the data. MIT, ASP, AVE, YuIV and VVM prepared the manuscript for submis-sion.

R E F E R E N C E S1. Patai Á., Solymosi N., Mohácsi L., Patai Á.V.

Indomethacin and diclofenac in the prevention of post-ERCP pancreatitis: a systematic review and meta-analysis of prospective controlled trials. Gastrointest Endosc. 2017 Jun;85(6):1144–1156.e1. doi: 10.1016/j.gie.2017.01.033

2. Dumonceau J.M., Andriulli A., Elmunzer B.J., Mariani A., Meister T., Deviere J., Marek T., Baron T.H., Hassan C., Testoni P.A., Kapral C.; European Society of Gastrointestinal Endoscopy. Prophylaxis of post-ERCP pancreatitis: European Society of Gastrointestinal Endoscopy (ESGE) Guideline - updated June 2014. Endoscopy. 2014 Sep;46(9):799–815. doi: 10.1055/s-0034-1377875

3. Zhang H., Cho J., Buxbaum J. Update on the Prevention of Post-ERCP Pancreatitis. Curr Treat Options Gastroenterol. 2018 Dec;16(4):428–440. doi: 10.1007/s11938-018-0194-y

4. Tenner S., Baillie J., DeWitt J., Vege S.S.; American College of Gastroenterology. American



College of Gastroenterology guideline: manage-ment of acute pancreatitis. Am J Gastroenterol. 2013 Sep;108(9):1400–15; 1416. doi: 10.1038/ajg.2013.218

5. Morales S.J., Sampath K., Gardner T.B. A Review of Prevention of Post-ERCP Pancreatitis. Gas-troenterol Hepatol (N Y). 2018 May;14(5):286–292. PMID: 29991936; PMCID: PMC6034611.

6. Mine T., Morizane T., Kawaguchi Y., Akashi R., Hanada K., Ito T., Kanno A., Kida M., Miyagawa H., Yamaguchi T., Mayumi T., Takeyama Y., Shimosegawa T. Clinical practice guideline for post-ERCP pancreatitis. J Gastroenterol. 2017 Sep;52(9):1013–1022. doi: 10.1007/s00535-017-1359-5

7. van Wanrooij R.L.J., van Hooft J.E. Prevention of post-ERCP pancreatitis: NSAID or pancreatic stent or both? Endosc Int Open. 2019 Jul;7(7):E869–E870. doi: 10.1055/a-0889-7796

8. Radadiya D., Devani K., Arora S., Chari-laou P., Brahmbhatt B., Young M., Reddy C. Peri-Procedural Aggressive Hydration for Post Endoscopic Retrograde Cholangiopancreatography

(ERCP) Pancreatitis Prophylaxsis: Meta-analysis of Randomized Controlled Trials. Pancreatology. 2019 Sep;19(6):819–827. doi: 10.1016/j.pan.2019.07.046

9. Maranki J., Yeaton P. Prevention of post-ERCP pancreatitis. Curr Gastroenterol Rep. 2013 Nov;15(11):352. doi: 10.1007/s11894-013-0352-2

10. Talukdar R. Complications of ERCP. Best Pract Res Clin Gastroenterol. 2016 Oct;30(5):793–805. doi: 10.1016/j.bpg.2016.10.007

11. Park T.Y., Oh H.C., Fogel E.L., Lehman G.A. Prevention of post-endoscopic retrograde cholangio-pancreatography pancreatitis with rectal non-steroidal anti-inflammatory drugs. Korean J Intern Med. 2020 May;35(3):535–543. doi: 10.3904/kjim.2020.069

12. Turovets M., Popov A., Mandrikov V., Vedenin Yu., Ekstrem A. Thoracic epidural anal-gesia for the prevention of post-ERCP pancreatitis: a randomized study of 491 cases // Archiv EuroMedica. 2020. Vol. 10. № 1. P. 69–75. doi: 10.35630/2199-885X/2020/10/18



CLINICAL SIGNIFICANCE OF LAMININ AND ELASTINE LEVELS IN CHILDREN WITH UNDIFFERENTIATED CONNECTIVE TISSUE DISEASE

A B S T R A C T — This research aims to study levels of laminin and elastine in 64 children with undifferentiated connective tissue disease (UCTD). All the children underwent clinical, laboratory and instrumental examination. It was found that changes in the levels of laminin and elastin are directly related to the severity of UCTD in children. Thus, clinical values of laminin and elastin levels can serve as additional criteria of UCTD severity. Using them, along with early detection of the phenotypic and visceral signs, helps prevent the development of severe forms of the disease.

K E Y W O R D S — undifferentiated connective tissue disease (UCTD), laminin, elastine, blood serum, severity criteria, children.

Nikolay Cherkasov , Yuliya Lutsenko


[email protected]

I N T R O D U C T I O NThe importance of detecting the undifferentiated

connective tissue disease is challenged by its high in-cidence (up to 80%) and ambiguous interpretation of phenotypic and visceral signs. The diagnostic criteria for the degree of its severity are proposed by research-ers from different countries [1, 2, 3, 4].

Evaluating the UCTD severity basing on phe-notypic and visceral manifestations has often resulted in uncertainty and frustration, thus urging the search of additional biochemical criteria, including such important non-collagen proteins as elastine, laminin, etc. They control interrelation between the elastic core and microfibrils [ 5, 6, 7]. Considering their sen-sitivity and extensive diagnostic range, their detec-tion may improve the evaluation of UCTD severity in children [8, 9]. All that justifies the urgency of the issue.

The aim of the research is to study clinical significance of laminin and elastine levels in children with UCTD.


http://dx.doi.org/10.35630/2199-885X/2021/11/2/P.1

C H A R A C T E R I S T I C O F T H E C H I L D R E N ; M E T H O D S O F R E S E A R C H

64 children aged 3 to 10 years with UCTD have been under observation. 27 of them had mild UCTD severity (the 1st group), 19 possessed medium UCTD severity (2nd group), and 18 suffered from severe UCTD (3rd group). The control group consisted of 18 conditionally healthy children.

UCTD diagnosis was performed basing on life record data, phenotypic and visceral signs, excluding genetic syndromes. Over 6 external phenotype data was analyzed. Ultrasonic scans of visceral organs were carried out.

The levels of elastine and laminin in blood serum were examined with the help of sandwich-type polarization fluoroimmunoassay, using the kits pro-duced by «Cloud-Clone Corp» (USA) to determine elastine quantifiable concentration (serial number HBE-337Hu No 4 DF58C0861) and laminin quan-tifiable concentration (serial number HBE082Hu No 7B578DDCCD). Data processing was performed with the methods of variable-based statistics (Statis-tika-10).

R E S U L T S A N D I N T E R P R E T A T I O NWe defined 6–7 phenotypic signs in the first

group. 65% of cases presented visceral signs of UCTD, including 13 cases of minor cardiac abnormalities (50%), 6 cases of mild pyelectasis (22,2%), 3 cases of nephroptosis (11,0%), and 3 cases of hypotension and cholecystis anomaly (11,0%). The levels of laminin and elastine in this group were close to normal range (p1 > 0,05; p2 >0,05).

Within the second group of children, 95,8% had 10–12 moderate phenotypic signs, early evidence of which was observed in 5 children (26,3%). The visceral signs were presented in 14 (72,8%) of minor cardiac abnormalities, 5 cases of pyelectasis (26,3%), 3 cases of nephroptosis (15,8%), 3 cases of gallblad-der volvulus (15,8%), and 5 cases with combination of signs (26,3%). The combination of phenotypic and visceral signs is 20% higher in the second group com-paring with the first one.

P E D I A T R I C S

https://orcid.org/0000-0001-7353-3615

https://orcid.org/0000-0002-7714-7612

http://dx.doi.org/10.35630/2199-885X/2021/11/2/P.1


The increase of elastine and laminin levels is statis-tically-valid (Table 1, p1< 0,01, p2< 0,05). The increase of proteins levels was associated with incidence of visceral signs combination (k1 = 0,72; k2= 0,61).

As for the third group, the infantile onset of UCTD (in the first years of life) dysplastic indicants in 7 children (36,7%) was interpreted as dysplastic march with early incipience. All the children under observa-tion showed 13–19 common phenotypic signs of mild and medium severity.

15 children under observation (83,4%) had combination of 15–17 phenotypic signs. Visceral signs were presented in 17 cases of minor cardiac abnormali-ties (95%), 7 cases of pyelectasis (38,8%), 4 cases of ne-phroptosis (22,2%), 3 cases of hydronephrosis (16,6%), 5 cases of gallbladder hypotension (26,7%), and 5 cases of gallbladder volvulus (26,7%). Comorbid syndromes (2,7) of renal and renocardial UCTD were present in 9 children (50,0%). Comorbidity of visceral signs in the third group was observed by 25% more of-ten than in the second group, and was associated with increase in laminin and elastine levels correspondingly (k1= 082; k2= 0,57).

Statistically significant difference (p1< 0,01, p2 <0,01) was observed between the levels of laminin and elastine in groups with mild, medium and severe UCTD (see Table 1). Consequently, we detected changes in laminin and elastine levels in direct depend-ence on UCTD severity in children. The determined values of these proteins’ levels may be used as addition-al criteria for evaluation of UCTD severity (Table 1).

Table 1. The levels of laminin and elastine in groups with mild, medium and severe UCTD

Note: * — reality in comparison of data of the 1st, 2nd and 3rd groups with control: (* >0,05, ** p<0,05, *** p<0,01; # — reality in comparison of data of the 1st and 2nd groups: # — p>0,05; ## — p<0,05

Data

GroupFirst(п=27)

Second(п=19)

Third(п=18)

Control(п=18)

laminin,(pg/ ml)

*, #47,2 ± 6,3

***56,3 ± 7,8

***62,3 ± 8,2 24,20 ± 7,1

elastine(ng/ml)

*, # 9,2 ± 2,5

**14,3 ± 1,7

***16,5± 1,4 7,2 ± 2, 2

reasonable evaluation criteria of UCTD severity. The use of phenotypic, visceral and biochemical indica-tors of undifferentiated connective tissue disease may contribute to timely and exact diagnosis. Moreover, it may help to start the treatment sooner.

R E F E R E N C E S1. Abbakumova L. N., Arsentyev V. G., Gnusaev

S. F. et al. Multifactorial and hereditary connective tissue disorders in children. Diagnostic algorithms. Management tactics. Russian guidelines. Pediatrist. 2016; N7 (2): P 5–39 (in Russ.).

2. Zueva T. V., Zhdanova T. V., Uraslina S. E. Comorbidity of renal and cardiac pathology. Medi-cal Bulletin of the North Caucasus. 2019; N 4(14): P711–715 (in Russ.).

3. Beighton P.,Paepe et al. International Nosology of Heritable Disorders of Connective Tissue – Berlin, 1988–29: 581 – 594. doi: 10.1002/ajmg.1320290316.

4. Аtarroyo M.,Tryggvason K.,Virtanen I. Laminin isoforms in tumor invasion, angiogenesis and metastasis. Semin.Cancer Biol. 2002. – N 12, P.197 – 207. doi: 10.1016/S1044-579X(02)00023-8.

5. Givant –Horwitz V. Davidson B.,Reich R. Laminin induced signaling in tumor cells. Cancer Lett 2005 N 223:P. 1–10. Lett 2005 N 223 P. 1–10.doi: 10.1016/j.canlet.2004.08.030.

6. Cologmato H., Yurchenco Р.D. Form and Function; The Laminin Family of Heter-otrimers. Developmental Dynamics. – 2000. – N 218: Р. 213- 234. doi:10.1002/(SICI)1097-0177(200006)218:2<213::AID-DVDY1>3.0.CO;2-R.

7. Lutsenko Yu. A., Cherkasov N. S., Davidova O.V., Ledyaev M.Ya., Makukhina L.P. et al. Clinical and instrumental assessment of forms and syndromes undifferentiated connective tissue dysplasia in children. Bulletin of the Volgograd state medi-cal University. 2019; N 3((71): P. 57–61 (in Russ.). doi:10.19163/1994-9480-2019-3(71)-58-61.

8. Mutalov A. G. Comorbid pathology in the practice of a pediatrician-features of diagnosis and manage-ment tactics. (Electronic resource). URL:https://medvestnik.ru/content/medarticles (access date: 4.12.2020) (in Russ.).

9. Mosca M., Tani C., Neri C., Baldini C., Bom-bardieri S. Undifferentiated connective tissue diseas-es (UCTD). Autoimmunity Reviews, No 6: 1, 2006, 1–4. https://doi.org/10.1016/j.autrev.2006.03.004

10. Laczik R, Soltesz P, Szodoray P, Szekanecz Z et al. Impaired endothelial function in patients with undifferentiated connective tissue disease: a follow-up study. Rheumatology, No 53: 11, 2014, 2035–2043, https://doi.org/10.1093/rheumatology/keu236

C O N C L U S I O NEarly evidence of visceral signs and their co-

morbidity are significant for UCTD diagnosis. It was found that changes in the levels of laminin and elastin are directly related to the severity of UCTD in children. Levels of laminin and elastine may serve as

P E D I A T R I C S


FUNCTIONAL PROPERTIES OF PLATELET IN CHILDREN WITH IRRITABLE BOWEL DISEASE

A B S T R A C T — The aim of the study was to evaluate platelet activation and aggregation in children with irritable bowel disease (IBD), as well as the effect of hyperbaric oxygenation on these processes. Platelet activation and aggregation in artificial shear flow were studied in 120 patients of both sexes aged 6 to 17 years with IBD. In pediatric patients with CD and UC, a significant increase in the activation and aggregation of platelets was revealed under shear flow conditions. The use of hyperbaric oxygen therapy leads to a decrease in the studied processes. It was found that IBD in children is characterized by significant changes in the functional properties of platelets (activation and aggregation processes).

K E Y W O R D S — children, irritable bowel disease, platelet, hyperbaric oxygen therapy (HBOT).

Alexandra Popovicheva , Andrew Martusevich ,Elvira Fedulova

Privolzhsky Research Medical University, Nizhny Novgorod, Russia

[email protected]

free environment) and their aggregation in children with IBD.

M A T E R I A L A N D M E T H O D SThe study used the blood samples of 120 patients

of both sexes aged 6 to 17 years, suffering from IBD. The study was approved by the Local Ethics Com-mittee of Privolzhsky Research Medical University. Informed consent to participate in the study was obtained from the parents of all children (or from the children themselves over the age of 15). The diagnosis of "IBD" was verified by the data of a comprehensive examination, which included clinical and laboratory data, as well as endoscopic examination of the intes-tinal mucosa with morphological analysis of biopsies. Blood sampling was performed at the time of hospi-talization of patients in the acute stage and at the end of treatment, before discharge. The treatment was car-ried out with the use of 5-aminosalicylic acid deriva-tives, glucocorticosteroids, immunosuppressants, and genetically engineered biological therapy. The results of the studies were compared with similar indicators of 35 conditionally healthy children of both sexes of the same age, who made up the control group.

Spontaneous (flow-induced) platelet aggregation was studied under conditions of artificial shear flow on a rheoscope designed according to the principle [4] in the modification [5]. The blood plasma was placed in a chamber of the device, in which a flow was created with a shear rate of 40 s-1. Discrete microphotography of the aggregation process was performed at an interval of 20 seconds after the start of aggregation for 400 seconds. The degree and speed of aggregation were determined by the number of aggregates (rel.units) after 400 s and 160 s, respec-tively, after the start of the process using a special program [6]. The determination of platelet activation in the artificial shear flow was carried out similarly to the above-described method. However, K3EDTA-stabilized blood plasma was used for this purpose. The determination of platelet aggregation under these conditions makes it possible to judge the presence of receptors on their membranes in high-affinity (the presence of platelet activation) or low-affinity (the absence of platelet activation) states. In the presence of platelet aggregation, its degree was estimated by the


I N T R O D U C T I O NInflammatory bowel diseases (IBD) are accom-

panied by quantitative, morphological and functional changes in platelets. The activated, hyperaggregable platelets are a key factor in enhancing the thrombo-genic potential and intestinal microinfarctions. In addition, platelets initiate an inflammatory phenotype in endothelial cells and white blood cells and promote increased inflammation by secreting numerous bio-logically active substances [1]. Therefore, it becomes clear that it is necessary to study first of all the activa-tion of platelets, which underlies the performance of their functions. To date, platelet activation is evaluated by measuring the concentration of circulating activa-tion markers (P-selectin, CD40, and GP53) not only in adult patients, but also in children [2, 3]. Platelet activation is determined by the mechanisms that cause calcium to enter platelets and lead to conformational changes in the GP IIb/IIIa receptor complex on the cell surface and, consequently, the final common path-way — platelet aggregation.

The aim of the study was to evaluate the activa-tion of platelets in an artificial shear flow (in a calcium-

P E D I A T R I C S

http://dx.doi.org/10.35630/2199-885X/2021/11/2/20

https://orcid.org/0000-0002-3717-2186

https://orcid.org/0000-0002-0818-5316

https://orcid.org/0000-0002-1774-0692

http://dx.doi.org/10.35630/2199-885X/2021/11/2/20


number of aggregates (rel.units) 400 s after the start of the aggregation process [7].

Statistical processing of the results was performed using variation statistics algorithms using Microsoft Excel 2007 and Statistica 6.1 for Windows.

R E S U L T SAs shown by the conducted studies, when the

EDTA blood was stabilized, spontaneous aggrega-tion of platelets in the control group under shear flow conditions was practically absent (Table 1). In children with IBD, the degree of platelet aggregation in these conditions exceeded the control values by 9 times, and continued to increase at discharge (Table 1).

Table 1. Activation of platelets in children with IBD (at 400 sec)

* р<0,05, ** р<0,001 — comparison to control

R E F E R E N C E S1. Lagrange J., Lacolley P., Wahl D., Peyrin-Bir-

oulet L., Regnault V. Shedding light on hemos-tasis in patients with inflammatory bowel diseases // Clin. Gastroenterol. Hepatol. – 2020. – S1542–3565 (20)30056-2.doi: 10.1016/j.cgh.2019.12.043

2. Matowicka-Karna J. Markers of inflammation, activation of blood platelets and coagulation disorders in inflammatory bowel diseases // Postepy Hig. Med. Dosw. (Online). – 2016. – Vol. 70. – P. 305–312. doi: 10.5604/17322693.1199305.

3. Ashkinazi V.I., Lebedev M.Y., Krestova E.I., Fedulova E.N., Shirokova N.Y., Tutina O.A.. Diagnostic value of leukocyte modulating serum activity factors in children with inflammatory bowel diseases // Pediatria. Journal them G.N. Speransky. – 2016. – Vol. 95, N 6. – P. 44–49.

4. Schmid-Schönbein H., von Gosen J., Heinich L., Klose H.J., Volger E. A counter-rotating "rheo-scope chamber" for the study of the microrheology of blood cell aggregation by microscopic observation and microphotometry // Microvasc. Res. – 1973. – Vol. 6, N 3. – P. 366–376. doi: 10.1016/0026-2862(73)90086-1.

5. Levin G.Ya., Modin A.P., Kudritsky S.Yu., Sosnina L.N. Method for determination of molondi-aldegide in blood. RF patent № 2278381.

6. Shagalova P.A., Sokolova E.S., Levin G.Ya., Zakharova A.Yu., Korotaev V.S., Kulikov G.V. Computer vision algorithms in the task of processing a series of medical images obtained from a micro-scope // Cloud of Science. – 2020. – Vol. 7. N 1. – P. 180–188.

7. Levin G.Ya., Egorikhina M.N. Method for deter-mining intravascular platelet activation. RF patent № 2416796.

Count of aggregates (rel. un.)control (n=35) 1,89±0,91

IBD (n=120)Before treatment 17,41±3,30 **After treatment 19,80±3,95 **

Count of aggregates (rel. un.)160 sec 400 sec

control (n=35) 20,03±0,96 15,28±1,09

IBD (n=120)Before treatment 25,60±0,92 ** 21,05±1,08 *After treatment 23,60±1,11 * 19,73±1,25 *

During hospitalization of children with IBD, there was also a significant increase in the degree and rate of spontaneous (flow-induced) platelet ag-gregation (by 1.4 times and 1.3 times, respectively) (Table 2). Platelet aggregation indicators remained sig-nificantly elevated in IBD and as a result of treatment. In addition, a positive correlation was found between the number of aggregates formed during platelet acti-vation and the same aggregation index during hospi-talization of patients with IBD (r = 0.50, p < 0.05).

Table 2. Spontaneous (flow-induced) aggregation of platelets in children with IBD

* р<0,05, ** р<0,001 — comparison to control

C O N C L U S I O NIBD in children is characterized by a significant

increase in platelet activation and aggregation. The use of HBOT in the treatment of IBD contributes to a significant reduction in these processes.

P E D I A T R I C S


CHARACTERISTICS OF GUT MICROBIOTA IN AUTOIMMUNE DISEASES OF THE THYROID GLAND AND THE METHODS OF ITS CORRECTION

A B S T R A C T — Gut microbiota is considered as a pathogenetic factor of various diseases nowadays. The patients with autoimmune diseases are known to suffer from dysbiosis. There are studies in the modern literature that demonstrate changes in the composition of the gut microbiota in case of thyroid dysfunction. This review examines a contemporary view of the gut microbiota, its role in the development of autoimmune diseases. We investigated the interaction between the thyroid gland and the gut microbiota, its species composition in hypo- and hyperthyroidism. Possible methods of correction, including the use of pre- and probiotics and transplantation of fecal microbiota have been demonstrated.

K E Y W O R D S — gut microbiota, autoimmune diseases, thyroid gland, autoimmune thyroiditis, hypothyroidism.

Nikolay Sturov1 , Sergey Popov1 ,Alina Kostenko1 , Grigory Roitberg2,3

1 RUDN University, Moscow2 N.I. Pirogov Russian National Research Medical University (RNRMU), Moscow3 “Medicinа” JSC, Moscow, Russia

[email protected]

Received 30 March 2021; Received in revised form 3 May 2021; Accepted 7 Mai 2021

to have a multifactorial etiology, including genetic, environmental factors, and food habits. Changes in the gut microbiota (GM) are one of the reasons for the development of AIDs. It has been proven that patients with AIT and GD have dysbiosis. Our review will con-sider the interaction between GM and the develop-ment of AIDs, the characteristics of GM composition in patients with TG’s dysfunction and methods of its correction.

R O L E O F G M I N T H E D E V E L O P M E N T O F A U T O I M M U N I T Y

The issue of the interaction between the com-position of human GM and AIDs is actively studied nowadays. The intestine is the main site of interaction between pathogenic bacteria, food antigens and nor-mal microflora. Epithelial cells, goblet cells producing mucous secretion, immune M-cells and lymphoid tissue are all the components of the intestinal barrier, the damage of which increases the host susceptibility to the effects of various agents [3]. The study of the intestinal wall structure showed similar changes in patients with type 1 diabetes and AIT, such as partial rarefaction of microvilli on the apical side of entero-cytes, a decrease in their height [4]. A change in the GM profile can damage transmembrane proteins re-sponsible for intestinal barrier integrity, such as ZO-1 and occludin [3]. The contact of intestinal immune cells with antigens and an increase in the intestinal permeability lead to inflammation and auto-aggression against the body's own cells.

Microbial imbalance is one of the pathogenetic factors in the development of AIDs. In dysbiosis, the ratio of regulatory Treg and inflammatory Th17 changes can also increase the susceptibility to patho-genic invasion [5]. Autoantibodies against the cell wall of Saccharomycecerevisiae have been found in patients with type 1 diabetes, rheumatoid arthritis (RA), systemic lupus erythematosus, antiphospholipid syndrome, and Crohn's disease [4]. Multiple sclerosis (MS) is an AID characterized by demyelination of nerve fibers. Analysis of the 16s ribosomal RNA gene showed the presence of dysbiosis in patients with MS. Colonization of Clostridium perfringens is associated

I N T R O D U C T I O NThe prevalence of autoimmune diseases (AIDs) of

the thyroid gland (TG), such as chronic autoimmune thyroiditis (AIT) and Graves' disease (GD) (diffuse toxic goiter), is estimated at about 5% [1]. AIT is a chronic inflammatory disease characterized by the production of specific autoantibodies against peroxi-dase (TPO-ab) and thyroglobulin (TG-ab) with the development of reduced thyroid function — hypothy-roidism (HT) [1]. AIT makes about 46% of the total TG’s pathology. According to the epidemiological data, the carrier state of TPO-ab is more common in the female population. There is also an increase in the incidence of TG’s dysfunction with age. TG’s hyperfunction is observed with GD and the main au-toantigen is the thyroid-stimulating hormone (TSH) receptor [2]. At present, AIDs of TG are known

T H E R A P Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/21

https://orcid.org/0000-0002-3138-8410

https://orcid.org/0000-0002-0567-4616

https://orcid.org/0000-0002-5729-4658

http://dx.doi.org/10.35630/2199-885X/2021/11/2/21


with a relapse of MS, and the produced toxins can cause microvascular complications leading to neuronal damage. Patients with severe dysbiosis and Sjogren's syndrome had higher disease activity and increased calprotectin level, which indicates the presence of an inflammatory process in the intestine. In the study of fecal microbiota in children with type 1 diabetes, representatives of the Blautia genus dominated, and their content positively correlated with the level of gly-cated hemoglobin and titers of antibodies to tyrosine phosphatase. The authors have suggested that it was the GM that influenced autoimmunity and may affect the development of type 1 diabetes [4]. C. aerofaciens may affect the pathogenesis of RA through various mechanisms, for example, by increasing the perme-ability of the intestinal wall, production of IL-17, and chemotaxis of neutrophils [6].

Thus, it can be assumed that changes in intestinal permeability and dysbiosis are the links in the patho-genesis of AIDs.

G M I N F L U E N C E O N T H Y R O I D F U N C T I O N . F E A T U R E S O F G M C O M P O S I T I O N I N A I D S O F T H E T H Y R O I D G L A N D

In recent years, the study of the Thyroid-Gut-Axis has been of increasing interest to researchers. The for-mation of metabolically active triiodothyronine (T3) occurs with the participation of enzymes — deiodi-nases, localized in many body tissues, including the intestinal wall. The authors have suggested that in view of the large surface of the GIT, the contribution of T3 activated by the deiodinases to the total body pool is substantial. GM metabolic by-products, especially short-chain fatty acids (SCFA), together with TG’s hormones, affect the differentiation of enterocytes and strengthen intercellular contacts in the intestinal wall [7]. GM participates in the metabolism of microele-ments, which are involved in the synthesis of hor-mones and the maintenance of normal TG function. With AIDs of the TG, there is a change in the levels of Lactobacillaceae and Bifidobacterium, the quantitative composition of which, in turn, positively correlates with the levels of selenium and zinc in the blood se-rum. It was suggested that disruption in the regulation of these microelements and GM composition could affect the course of AIT and GD [8].

There is evidence that microorganisms that are structurally homologous to human proteins through the mechanisms of molecular mimicry can cause AIDs of the TG. E.P. Kiseleva et al. have found Bifido-bacteriumadolescentis, B. Longum, B. bifidum, and Lactobacillus plantarum on cells surfaces; these are the components that compete with human TPO-ab

and TG-ab. The dependence between the presence of antibodies to B. bifidumand L. plantarum, and TPO-abwas also obtained [9]. Gram-positive bac-teria, including strains of Lactobacillus rhamnosus, can activate the transcription factor NF-κB, which controls the expression of genes responsible for the immune response, either directly or through cytokines. In the study by Jiang W. et al. in patients with GD in GM analysis, an increase in the strains of Lactobacillus and Bacteroides was revealed, and in some patients, increased titers of TPO-ab were observed [10]. The authors have assumed that hormone imbalance in GD and changes in GM towards the predominance of Lactobacillus, which had similar amino acid sequences with TPO-ab and TG-ab led to the induction of proinflammatory signaling pathways with the develop-ment of autoimmunity.

GM can affect the predisposition to the devel-opment of HT in mouse models. As far back as in 1988, Penhale W.J. et al. demonstrated that injection of an antibiotic by sterile rats followed by injection of homogenized intestinal contents from rats raised under normal conditions increased their autoimmune susceptibility [10].

It is known that AIDs of the TG are character-ized by dysbiosis. Some studies point out that there is a decrease in microbial diversity; others, on the contrary, note the small intestinal bacterial overgrowth syndrome (SIBO). The research of Yan HX. et al. showed that the species composition of GM was less diverse in patients with HT, and Phascolarctobacte-rium predominated [11]. In the group with GD, the diversity of microbial strains also decreased and there were changes in the GM profile in the form of an increase in the number of Bacilli, Prevotella, Lacto-bacillales, Megamonas, Veillonella and a decrease in Rikenellaceae, Ruminococcus, and Alistipescompared with healthy people [12]. In HT, there is a decrease in the motor function of the digestive system due to the accumulation of mucopolysaccharides and the development of the intestinal wall edema. Decreased GIT vermicular movement is one of the factors in the development of SIBO. The study involving 1809 peo-ple demonstrated that the most important factors in the development of SIBO were immunosuppression, conditions after gastric/intestinal resection and HT [13]. Lauritano E.C. et al. have found that about 54% of patients with HT suffered from SIBO [14]. Zhao F. et al. have found bacterial overgrowth and differences in the GM composition of patients suffering from HT and healthy individuals in the form of increased genera Blautia, Romboutsia, EubacteriumRoseburia, Rumi-nococcus, Fusicatenibacter, Dorea and Eubacterium and decreased levels of Fecalibacterium, Bacteroides,

T H E R A P Y


Prevotella [15]. The change in GM correlated with clinical parameters, which could be actively used for the diagnosis of diseases. Another study also demon-strated a change in GM with a predominance of Shig-ella, Escherichia coli, and Parasutterella and a decrease in the content of Prevotella and Dialister [16].

It is known that GM affects the absorption of drugs. Impaired absorption of levothyroxine (LT4) can be caused by a change in the GM composition due to concomitant diseases of the GIT. There is evidence in the literature that the prevalence of coeliac disease- AID, which is characterized by gluten intolerance, with AIT, according to various sources, ranges from 5 to 24%. In the study by Bibbò S. et al. in patients with AIT and coeliac disease, a decrease in the level of Bifidobacterium was observed [17]. Virili C. et al. obtained that patients with isolated HT achieved nor-malization of the TSH level (on average, 1.02 mU/l) against the background of LT4 therapy at a daily dose of 1.31 μg/kg, while in patients with HT and coeliac disease during the same period of treatment with similar doses, higher TSH values (4.20 mU/l) were observed [18]. It was found that when correcting HT with concomitant coeliac disease, it was neces-sary to increase the daily dose of LT4, on average, to 1.96 μg/kg [19]. Patients infected with Helicobacter pylori also required large doses of LT4 to achieve com-pensation for HT.

C O N C L U S I O N GM plays an essential role in the human body,

performing many functions: immunomodulatory, biosynthetic, protective, etc. With the advent of new non-culture-based research methods, it became pos-sible to study the qualitative and quantitative composi-tion of GM in various diseases. A change in the GM profile can be considered as one of the factors in the development of AIDs. The patients with autoimmune TG’s dysfunction are known to suffer from dysbiosis. Evaluating the relevant literature data, it can be as-sumed that there is an interaction between hormone imbalance and GM. There is no doubt concerning the necessity for further clinical studies to identify the characteristics of the GM composition in AIDs of the TG. The detection of differences between normal and transformed GM can be considered as a diagnostic criterion for TG’s diseases. Further study of GM cor-rection methods will help to determine new strategies for the treatment and prevention of TG’s diseases.

R E F E R E N C E S1. Tomer Y. Mechanisms of autoimmune thyroid dis-

eases: from genetics to epigenetics. Annu Rev Pathol. 2014;9:147–56. https://doi.org/10.1146/annurev-pathol-012513-104713

2. Rayman MP. Multiple nutritional factors and thyroid disease, with particular reference to autoimmune thyroid disease. Proc Nutr Soc. 2019;78(1):34–44. https://doi.org/10.1017/S0029665118001192

3. Yamamoto EA, Jørgensen TN. Relationships Between Vitamin D, Gut Microbiome, and Systemic Autoimmunity. Front Immunol. 2020;10:3141. ht-tps://doi.org/10.3389/fimmu.2019.03141

4. Sasso FC, Carbonara O, Torella R, et al. Ultrastructural changes in enterocytes in subjects with Hashimoto's thyroiditis. Gut. 2004;53(12):1878–80. https://doi.org/10.1136/gut.2004.047498

5. Covelli D, Ludgate M. The thyroid, the eyes and the gut: a possible connection. J Endocrinol Invest. 2017;40(6):567–576. https://doi.org/10.1007/s40618-016-0594-6

6. Vázquez NM, Ruiz-Limón P, Moreno-Indias I, et al. Expansion of Rare and Harmful Lineages is Associated with Established Rheumatoid Arthritis. J. Clin Med. 2020;9:1044. https://doi.org/10.3390/jcm9041044

7. Fröhlich E, Wahl R. Microbiota and Thyroid Interaction in Health and Disease. Trends Endo-crinol Metab. 2019;30(8):479–490. https://doi.org/10.1016/j.tem.2019.05.008

8. Knezevic J, Starchl C, TmavaBerisha A, Amrein K. Thyroid-Gut-Axis: How Does the Microbiota Influence Thyroid Function? Nutri-ents. 2020;12(6):1769. https://doi.org/10.3390/nu12061769

9. KiselevaEP, MikhailopuloKI, SviridovOV, et al. The role of components of Bifidobacterium and Lactobacillus in pathogenesis and serologic diagnosis of autoimmune thyroid diseases. Benef Mi-crobes. 2011;2(2):139–54. https://doi.org/10.3920/BM2010.0011

10. Penhale WJ, Young PR. The influence of the normal microbial flora on the susceptibility of rats to experimental autoimmune thyroiditis. Clin Exp Im-munol. 1988;72:288–292.

11. Yan HX, An WC, Chen F, et al. Intestinal micro-biota changes in Graves' disease: a prospective clinical study. BiosciRep. 2020;40(9):BSR20191242. https://doi.org/10.1042/BSR20191242

12. Yaylali O, Kirac S, Yilmaz M, et al. Does hypothyroidism affect gastrointestinal motility? Gas-troenterol Res Pract. 2009;2009:529802. https://doi.org/10.1155/2009/529802

13. Brechmann T, Sperlbaum A, Schmiegel W. Levothyroxine therapy and impaired clearance are the strongest contributors to small intestinal bacterial overgrowth: Results of a retrospective cohort study. World J Gastroenterol. 2017;23(5):842-852. https://doi.org/10.3748/wjg.v23.i5.842

14. Lauritano EC, Bilotta AL, Gabrielli M, et al. Association between hypothyroidism and small intestinal bacterial overgrowth. J ClinEndocrinolMe-tab. 2007;92(11):4180–4. https://doi.org/10.1210/jc.2007-0606

T H E R A P Y


15. Zhao F, Feng J, Li J, et al. Alterations of the Gut Microbiota in Hashimoto’s Thyroiditis Patients. Thy-roid. 2018;28(2):175–86. https://doi.org/10.1089/thy.2017.0395

16. Ishaq HM, Mohammad IS, Guo H, et al. Mo-lecular estimation of alteration in intestinal microbial composition in Hashimoto's thyroiditis patients. Biomed Pharmacother. 2017;95:865–874. https://doi.org/10.1016/j.biopha.2017.08.101

17. Bibbò S, Abbondio M, Sau R, et al. Fecal Microbiota Signatures in Celiac Disease Patients With Poly-Autoimmunity. Front Cell Infect Microbiol. 2020;10:349. https://doi.org/10.3389/fcimb.2020.00349

18. Virili C, Bassotti G, Santaguida MG, et al. Atypical celiac disease as cause of increased need for thyroxine: a systematic study. J ClinEndocrinolMe-tab. 2012;97(3):E419–22. https://doi.org/10.1210/jc.2011-1851

T H E R A P Y


BISMUT-INDUCED ONSET OF ACUTE TUBULOINTERSTICIAL NEPHRITIS: А СLINICAL CASE

A B S T R A C T — This article presents a clinical case of drug-induced acute TIN that developed in response to self-administered intake of bismuth tripotassium dicitrate. This clinical case was characterized by an acute onset, presence of urinary syndrome (low specific gravity of the urine and proteinuria), occurrence of azotemia in the setting of preserved diuresis, lack of hyperkaliemia, and quick and complete restoration of renal functioning after the discontinuation of a bismuth-containing drug. It can be suggested that the presented clinical case will allow for a better understanding of the causes and clinical features of acute TIN and the choice of the management tactics for patients with this pathology.

K E Y W O R D S — tubulointerstitial nephritis (TIN), medicinal nephritis, acute kidney damage, chronic kidney disease, bismuth trisalium citrate.

Maria Mikhailova , Olga Poselyugina

Tver State Medical University, Tver, Russia

[email protected]


B A C K G R O U N DDrug-induced renal injury is one of the most

acute problems in modern nephrology that is caused by the development of tubulointerstitial nephritis (TIN). As a rule, acute TIN is the main cause of unspecified kidney injury in patients with preserved diuresis and normal kidney sizes [1–5, 8]. TIN is an inflammatory condition of renal interstitium and tubules that is associated with the development of acute renal injury in 10–25% of cases. In 20–40% of cases, it leads to the development of chronic kidney disease (CKD). One of the drugs, which administra-tion can be complicated by the development of TIN, is bismuth tripotassium dicitrate that exerts anti-ulcerous and anti-inflammatory effects combined with bactericidal activity against Helicobacter pylori. This drug is widely used in the treatment of ulcerous disease and chronic gastroduodenitis in the exacerba-tion phase [6–13]. This drug contains bismuth oxide that is primarily eliminated by kidneys. It can be suggested that the expansion of the knowledge on the peculiarities of this pathology can contribute to its timely diagnostics.

The study aimed to describe a clinical case of drug-induced acute TIN that developed in response to intake of bismuth tripotassium dicitrate and expand the doctors’ knowledge about this pathology for the prevention, timely diagnostics, and treatment of acute TIN.

M A T E R I A L S A N D M E T H O D SThe authors reviewed medical publications on

clinical recommendations on drug-induced TIN for the past decade and analyzed the medical history of a patient from the Nephrology Department of the Tver Regional Clinical Hospital (Russia).

C A S E D E S C R I P T I O NPatient K., 56 years old, female, was admitted

to Nephrology department of the tertiary hospital complaining about pains in the epigastrium, feeling of abdominal borborygmi and bloating, loose stool up to 2–3 times per day, bilateral lumbar pain, and nausea.

Antibiotics were not prescribed in this case, because the patient self-medicated herself recognizing her symptoms as gastritis. She decided to consult a doctor only after taking the drug during 4 days, when her condition began to deteriorate.

The patient said that the condition developed quickly. The symptoms intensified within several days. The patient began to take bismuth tripotassium dicitrate 2 tablets twice a day without prescription. On day 4 after the beginning of the drug intake the patient developed face edema. The daily urine output decreased. During the examination at the local clinics, biochemical blood assay showed elevated levels of creatinine to 889.5 µmol/L and BUN to 21.6 µmol/L. The specified condition was defined as acute renal injury developed on day 4 after the drug intake. On day 6, the patient was hospitalized.

The patient’s condition was satisfactory at admis-sion to the hospital. Face edema and pastosity of the lower third ankles and feet were observed. The pulse was 80, rhythmic. BP was 16090 mmHg. The abdomen is soft, slightly painful in the epigastrium. The kidneys were not palpable. The diuresis was 1.0 L.

The clinical blood analysis revealed light normo-chromal anemia (hemoglobin 105 g/L), anisocytosis, light thrombocytopenia (167×109/L), eosinopenia,

T H E R A P Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/22

https://orcid.org/0000-0001-5775-9868

http://dx.doi.org/10.35630/2199-885X/2021/11/2/22


ESR acceleration to 27 mm/h. Common urine analysis showed urine low specific gravity (1008), insignifi-cant proteinuria (to 0.06 g/L), a large amount of flat epithelium, and 2–4 leukocytes in FOV. Biochemi-cal blood assay at admission showed hyperazotemia (creatinine — 850 µm/L, BUN — 15 mmol/L). Ultrasonic investigation revealed diffuse alterations in the renal parenchyma with an increase in their size. ECG registered sinus bradycardia, horizontal electri-cal axis of heart, and signs of left ventricular overload. Esophagogastroduodenoscopy revealed esophagitis, mixed gastritis, duodenogastric reflux, and indirect signs of cholecystopancreatitis. EchoCG protocol showed insignificant hypertrophy of the basal area of the interventricular septum, left ventricular diastolic dysfunction, and preserved ejection fraction. Gastro-enterologist and nephrologist examined the patient and evaluated the necessity of urgent hemodialysis. However, considering preserved diuresis, positive dynamics of creatinine levels decrease in the blood, and satisfactory condition, conservative management of the patient was chosen.

Clinical diagnosis was made based on the results of laboratory-instrumental tests. The primary diagno-sis was drug-induced acute tubulointerstitial nephri-tis developed after intake of bismuth tripotassium dicitrate. Complications: moderate acute renal injury. Comorbid diagnosis: gastro-esophageal reflux disease: I degree esophagitis. Chronic gastroduodenitis in the exacerbation phase. Fatty hepatosis. Pancreatic lipoma-tosis. Hypertensive disease, II degree, high risk.

In the Nephrology Department, the patient re-ceived conservative therapy that included discontinu-ation of bismuth tripotassium dicitrate, indication of euphylline infusions (2.4% — 10.0 i/v drop infusion BID, daily), omeprazole (40 g i/v drop infusion before dexamethasone infusion), dexamethasone (16 mg i/v drop infusion daily with a gradual dose decrease to complete discontinuation), prednisolone (20 mg per os starting a week after the beginning of treatment at hospital), furosemide (40–60 mg i/v). During the therapy, the patient’s condition improved and positive dynamics of laboratory parameters were observed.

At the patient’s discharge a significant decrease in the levels of creatinine (to 150 µmol/L) and BUN (to 11 µmol/L) were observed. Clinical blood analysis showed normalization of hemoglobin, erythrocyte, and platelet levels, and a decrease of ERS to 16 mm/h. A common urine test revealed insignificant proteinuria (0.03 g/L). For outpatient therapy, it was recommend-ed to take antiaggregants (pentoxifylline) 100 mg BID, per os, for 1 month. 1 month after the discharge, levels of creatinine and BUN were normalized.

D I S C U S S I O NThe presented clinical case included the major

manifestations of drug-induced acute TIN compli-cated by acute kidney injury developed in response to self-medication of bismuth tripotassium dicitrate. The clinical features of this case included acute onset (4 days after the self-medication), lack of oligo-anuria, presence of urinary syndrome (low specific gravity of the urine and proteinuria), increase in the level of creatinine in the blood with preserved diuresis, lack of hyperkaliemia, and quick and complete restora-tion of renal functioning after discontinuation of the bismuth-containing drug [10–13].

It can be suggested that the presented clinical case will allow better understanding of the causes and clini-cal features of acute TIN and the choice of manage-ment tactics for patients with this pathology.

R E F E R E N C E S1. Alyaev Yu. G., Glybochko P. V., Pushkar D. Yu.

// Clinical guidelines for the diagnosis and treatment of acute renal failure // Russian Society of Urologists. - 2014.

2. Association of general practitioners (family doctors) of the Russian Federation // Acute and chronic tubu-lointerstitial nephritis // clinical guidelines for general practitioners. – 2014.

3. Dyadyk A.I., Kugler T.E. Medicinal acute tubulointerstitial nephritis. // Almanac of Clinical Medicine. - 2017.– No. 45 (7). – p. 586–98.

4. Bomback A.S., Markowitz G.S. Increased prevalence of acute interstitial nephritis: more disease or simply more detection? Nephrol Dial Transplant. 2013; 28(1):16–18. DOI: 10.1093/ndt/gfs318 DOI: 10.1093/ndt/gfs318

5. Biruh T. Workeneh, MahendraAgraharkar, Rajiv Gupta // Acute Kidney Injury. – 2018.

6. Nast C. C. Medication-induced interstitial nephritis in the 21st century. // Adv Chronic Kidney Dis. 2017.–24(2). – р. 72–9. DOI: 10.1053/j.ackd.2016.11.016

7. Oyama S, Hosohata K, Inada A, Niinomi I, Mori Y, Yamaguchi Y, Uchida M, Iwanaga K Drug-induced tubulointerstitial nephritis in a retro-spective study using spontaneous reporting system database. Therapeutics and Clinical Risk Management. Vol.14 //2018. p. 1599-1604, DOI: 10.2147/TCRM.S168696

8. Goicoechea M, Rivera F, Lopez-Gomez JM; Spanish Registry of Glomerulonephritis. Increased prevalence of acute tubulointerstitial nephritis. Nephrol Dial Transplant. 2013;28(1):112–115 DOI: 10.1093/ndt/gfs143

9. Toto RD. Acute tubulointerstitial nephritis. Am J Med Sci. 1990;299(6):392–410. DOI: 10.1097/00000441-199006000-00007

T H E R A P Y


10. Gabow PA, Lacher JW, Neff TA. Tubulointersti-tial and glomerular nephritis associated with rifampin. Report of a case. JAMA. 1976;235(23):2517–2518. DOI: 10.1001/jama.235.23.2517

11. Wilson GJ, Kark AL, Francis LP, Hoy W, Healy HG, Mallett AJ. The increasing rates of acute interstitial nephritis in Australia: a single centre case series. BMC Nephrol. 2017;18(1):329. DOI: 10.1186/s12882-017-0747-7

12. Rossert J. Drug-induced acute interstitial ne-phritis. Kidney Int. 2001;60(2):804–817. DOI: 10.1046/j.1523-1755.2001.060002804.x

13. Saeki T, Nishi S, Imai N, et al. Clinicopatho-logical characteristics of patients with igg4-related tubulointerstitial nephritis. Kidney Int. 2010;78(10): 1016–1023. DOI: 10.1038/ki.2010.271

T H E R A P Y


EXPRESS METHOD FOR TREATING NEGATIVE EMOTIONAL STATES IN DENTISTS

A B S T R A C T — This paper is aimed to examine the prevalence of such negative emotional states as tension, anxiety and depression among students and dentists (n=80). Herein, we developed the "Method for treating negative emotional states" and analyzed its effectiveness. The data of questionnaire survey and determination of adaptive potential were assessed according to R.M. Baevsky. It was revealed that such parameter as decrease in the heart rate by 7 beats per minute could be used as a criterion to evaluate the effectiveness of treating negative emotional states in medical professionals. It has been proved that our original method is effective in managing negative emotional states, thus maintaining working ability, professional longevity and mental health among specialists and future doctors.

K E Y W O R D S — tension, anxiety, depression, pulse, adaptive potential, treatment of negative emotional states.

Petr Petrov , Sergey Averyanov , Sergey Lazarev, Ilgiz Iskhakov ,Adel Isaeva

Bashkir State Medical University, Ufa

[email protected]


R E L E V A N C EMedicine, in particular, dentistry is a type of hu-

man activity where negative emotions prevail, affecting individual’s physical and psychological well-being [5]. Dentists in general are characterized by a low level of health and quality of life, high morbidity, one of the reasons for which is neuro-emotional overload and high levels of stress [3, 12].

Initially, the concept of stress originating in physiology characterized a non-specific reaction of the body in response to any adverse effect known as general adaptation syndrome. In reference studies of stress development among dentists, it is noted that several reasons could be distinguished in the activities of these specialists that allow them to be attributed to a risk group: constant competition, hard and painstaking work that is performed in a limited space of the oral cavity, etc.

These problems cause a high incidence of cardio-vascular diseases, ulcers, colitis, neuromuscular pain, eye strain, and also affect family relationships, causing alcoholism, drug addiction, mental depression and

even suicide [13]. Prolonged occupational stress, in turn, leads to the emergence of burnout syndrome [8]. Emotional burnout is considered as a breakdown in adaptation system developing as a result of prolonged work stress caused by exposure to various stressors, in-cluding physical, chemical, biological, social, and even one's own thoughts. It is also believed that stress and burnout are relatively independent phenomena. Thus, the key difference between burnout and stress is the causal factors. Stress occurs in a myriad of situations (e.g., war, natural disasters, illness, unemployment, various situations at work, etc.), while burnout mani-fests itself in connection with professional activities [6, 11]. Stress may not necessarily cause burnout, and conversely, stress can have a mobilizing effect on the human body.

Scientists have found that the practice activ-ity of dentists at the age of 41–55 is characterized by predominance of negative emotions, maximum professional tension of personal adaptive resources that contribute to the formation of psychosomatic pathology in this contingent. At the same time, most of the dentists (64.3%) belong to the group of doctors who confirm the presence of psycho-emotional stress, 20.9% of which have signs of clear depletion of adapta-tion mechanisms [8].

When assessing the health condition in clini-cal practice, adaptive potential (AP) of circulatory system could serve as one of the effective health indicators, since cardiovascular system represent the primary adaptive mechanism of the whole organism that reacts in response to external conditions. It acts as a regulator of internal environment of the body, maintaining adequate blood supply and homeosta-sis of organs and tissues [7]. Considering the above mentioned evidence, we have chosen the well-known assessment of AP of circulatory system proposed by R.M. Baevsky [2].

Nowadays, there are many approaches for elimi-nating the breakdown of adaptation mechanisms such as psychotherapy, muscle relaxation, phyto- and drug therapy, etc. The majority of these therapeutic meth-ods have one drawback: such treatment should be car-ried out for a long time, months and years, sometimes throughout life.

Therefore, it is of particular importance to de-velop short-term and adequate psychological meth-ods aimed to prevent specific forms of psychological

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/23

https://orcid.org/0000-0003-1827-1629

https://orcid.org/0000-0002-6177-0573

https://orcid.org/0000-0002-7561-9145

http://dx.doi.org/10.35630/2199-885X/2021/11/2/23


tension and preserve professional longevity and mental health of specialists [1].

Aiming to solve the indicated problem, we have developed an express method to cope with such emotional states as tension, anxiety and depression and received a patent for this invention. The proposed method deals with embracing negative emotions via activation of the information processing system. It is effective, convenient, and most importantly, the thera-peutic time does not exceed 3–5 minutes.

Aim of the research: to evaluate the effectiveness of the author's method in treating emotional states in the study group.

The object of the research includes negative emotional states and its manifesta-tions in the practice of dentists and students of the dental faculty.

The subject of the research is the breakdown of AP in the study group and its nor-malization by application of the author's method.

R E S E A R C H O B J E C T I V E S1. To analyse the prevalence of emotional states in

the study group.2. Determine the AP of cardiovascular system in

the study group according to R.М. Baevsky.3. Eliminate the revealed emotional states using

the author's "Method of treating emotional states".

R E S E A R C H M A T E R I A L We examined 80 volunteers aged 21 to 64 years

old, with mean age 34.16±0.97 years; 39 men and 41 women. 25% of examined volunteers were students of the Dental Faculty of Bashkir State Medical University (BSMU), 10% were orthodontists, 65% were students of the Department of Prosthetic Dentistry and Maxil-lofacial Surgery at the Institute of Additional Profes-sional Education of BSMU.

R E S E A R C H M E T H O D STesting, measurement of pulse, systolic and diasto-

lic blood pressure was carried out twice in the examined volunteers: at rest while sitting in a chair (initial AP), and immediately after the intervention by the proposed author's method. It includes special actions with macro-saccades of eyes revealed during their smooth rotation and verbalized localization in the body of negative emo-tions [9]. For the objectivity of the experimental results, the volunteers received the treatment once.

Recognition of the functional state of volunteers was carried out on the basis of the test by V.V. Libinykh

and A.V. Libinykh modified by A.F. Yermoshina called /Constructive drawing of a person from geometric figures [4] and the questionnaire proposed by us, as well as the analysis of AP results before and after treatment.

AP was determined by the formula:

AP = (0.0011*HR) + (0.014*SBP) + (0.008*DBP) + (0.009*BW) – (0.009*H) + (0.014*A) – 0.27,

where HR is the pulse rate per minute, SBP is the level of systolic blood pressure and DBP — diastolic blood pressure, BW — body weight, H — height and A — age [2].

For statistical processing of the obtained results, MS Excel 10.0 software was used. The calculation was carried out by nonparametric method χ2 — Pearson's criterion with arithmetic mean values (M ± m).

R E S E A R C H R E S U L T SAnthropometric characteristics among the

subjects were as follows. The body height in fe-males ranged from 152 to 178 cm, on average was 163.29±0.85 cm, while in males ranged from 165 to 202 cm, on average — 176.00±1.26 cm. The body weight in women varied from 44 to 85 kg, the average was 62.05 ± 2.06 kg, while in men the body weight was much higher ranging from 60 to 114 kg, the average was 80.23 ± 1.94 kg.

The mean SBP at rest and after treatment was 124.31±21.15 and 123.73±17.58 mm Hg, while DBP, respectively, was 77, 72±10.9 and 80.36±13.58 mm Hg. HR before and after exposure was significantly different (χ2 = 0.000): before — 79, 51±17.41 beats per minute, after — 72, 35±11.16. The difference was approximately 7 beats per minute [10]. Therefore, decrease in the heart rate could be used in further research as evidence of the effectiveness in treating negative emotional states.

Consequently, all examined volunteers were di-vided into 4 groups according to the presence of nega-tive emotional states: I — those experiencing tension; II — anxiety; III — depression; IV — calmness.

According to the AP values, the functional state of the volunteers was determined and divided into the following groups:

I. AP below 2.6 corresponding to satisfactory adaptation of circulatory system;

II. AP between 2.6 and 3.09 corresponding to tension of adaptation mechanisms (practically healthy according to L.L. Miller [7]);

III. AP range from 3.10 to 3.49 corresponding to poor adaptation;

IV. AP of 3.5 and higher corresponds to adapta-tion failure.

D E N T I S T R Y


Following the therapeutic intervention by the proposed author's method, the total number of volun-teers experiencing negative emotional states decreased from 46.25% to 30.0% and was distributed as follows (Table 1).

Table 1. Distribution of emotional states among the volunteers before and after intervention (n=80, χ2=0,033)

Table 2. Distribution of AP before and after therapeutic intervention into the following groups (n=80, χ2=0,000)

of the respondents from the group II improved their indicators and moved to the group I with satisfactory adaptation. Meanwhile, the remaining 73.68% of cases from the second group remained the same. 22.22% of respondents from group III moved to group II, 7.69%

According to the interventionEmotion

Totaltension anxiety depression calmness

before 31,25% 15,0% 5,0% 48,75% 100%after 21,25% 5,0% 3,75% 70,0% 100%

Group of AP Before therapeutic interventionAfter therapeutic interventionI II III IV

I 50,0% 87,5% 12,5% – –II 23,75% 26,32% 73,68% – –III 22,5% 5,5% 22,22% 72,22% –IV 3,75% – – 33,33% 66,66%

At the same time, there were no significant dif-ferences between men and women. The portion of respondents experiencing anxiety decreased most of all by approximately 66.7%, those experiencing tension — by 32.0%. The number of volunteers experiencing depression decreased by almost 25.0%, while those feeling calmness increased by 30.35%.

Depending on the results of AP before and after therapeutic intervention, all volunteers were distrib-uted into the following groups (Table 2).

Before the therapeutic intervention, half of the volunteers were included in the group I according to the AP values, where the adaptation of circulatory system was satisfactory. The rest of the respondents were distributed as follows: 23.75% of the respondents entered the II and 22.5% — III group, corresponding to tension and unsatisfactory adaptation. The group IV was the smallest one and included those experienc-ing a breakdown in adaptation.

After the therapeutic intervention, the following results were obtained: a large proportion of the studied group I (87.5%) remained in the same group charac-terized by satisfactory adaptation with sufficiently high functional capabilities of the organism. The remaining 12.5% of the respondents from the first group experi-enced stress and moved to the second group. 26.32%

— to group I, thereby improving their AP indicators. 33.33% of respondents from group IV with "failure of adaptation" moved to the III group with unsatisfactory adaptation, and in terms of the numerical AP indica-tors improving their condition.

To conclude, a single treatment of negative emotional states with the proposed author's method resulted in the improvement of adaptive capabilities in 13.75% of respondents. At the same time, there were no cases of deteriorated AP parameters after the

therapeutic intervention as compared to the baseline levels.

C O N C L U S I O N1. Negative emotional states such as tension, anxi-

ety, depression are widespread among dentists.2. A statistically significantly specific effect of

our method on the body functional status has been confirmed.

3. A decrease in the heart rate in the adult popula-tion by 7 beats per minute can be used in scientific research as a reliable criterion for the effectiveness of treatment of emotional states.

4. The proposed therapeutic intervention method is effective in treatment of such negative emotional states as tension, anxiety and depression. It enables to

D E N T I S T R Y


maintain professional longevity and mental health of dental specialists and future doctors.

R E F E R E N C E S1. Bagriy M.A. Features of the development of profes-

sional stress in doctors of different specializations: abstract of thesis for a candidate of psychological sciences. – Moscow, 2009. – 29 p.

2. Baevsky R. M., Berseneva A. P. Assessment of the adaptive capacities of the organism and the risk of de-veloping diseases. - Moscow: Medicine, 1997. – 235 p.

3. Bezdetko G.I. Improvement of organizational forms of medical care in prosthetic dentistry on an outpa-tient basis: abstract of thesis for a candidate of medical sciences. – Moscow, 2019. – 25 p.

4. Ermoshin A.F. The geometry of experiences. Con-structive drawing of a person in psychotherapeutic practice. – M.: Neolit, 2017. – 288 p.

5. Ilyin E.P. Emotions and feelings: a tutorial / EP Ilyin. – 2nd ed., Rev. and add. – SPb [and others]: Peter, 2011. – 782 p.

6. Kazarina L.N., Vdovina LV. Psychosomatic dis-eases in dentistry // Review. Dentistry. – No. 1 (72). – 2011. – pp. 24–27.

7. Miller L.L. Sports medicine: textbook. – M .: "Hu-man", 2015. – 184 p.

8. Nefedov O.V. Physiological and hygienic character-istics of factors shaping the health of dentists: abstract dis. ... a candidate of medical sciences. – Moscow, 2017. – 24 p.

9. Patent No. 2486867 RF. A method of treating negative emotional states. / P.I. Petrov. – No. 2012123060, app. 06/04/12; publ. 10.07.13, Bul. No. 19. https://yandex.ru/patents/doc/RU2486867C1_20130710 (date of reference 03/08/2021).

10. Porges S. W. The polyvagal theory: neurophysiologi-cal foundations of emotions, attachment, communica-tion, and self-regulation (Norton Series on Interper-sonal Neurobiology). – WW Norton & Company, 2011.

11. Syurtukova E. Yu. Organizational and personal determinants of mental burnout in the activities of teachers of preschool educational institutions: abstract of thesis for a candidate of psychological sciences. – Yaroslavl, 2014 . – 27 p.

12. Besalat Hussain, Fatima Mushtaq, Asma Saher Ansari. Awareness of Role of Dentist–Patient Co–operation in Attainment of Ideal Dental Treat-ment among Dental Interns at DUHS Karachi //International Journal of Dental Sciences and Research. 2017. – Vol. 5. – No. 5. – Р. 116–121. doi:10.12691/ijdsr-5-5-1

13. Randy Lang, DDS, D. Ortho. Stress In Dentist-ry– It Could Kill You! [Electronic resource] /Sep-tember, No 1, – 2007: website. – URL: https://www.oralhealthgroup.com0/features/stress–in–dentistry–it–could–kill–you/ (date of reference 08.03.2021).

D E N T I S T R Y


IMMUNOLOGICAL ASSESSMENT OF THE STRESS RESPONSE IN PATIENTS WITH INFLAMMATORY POSTPROTHETIC COMPLICATIONS

A B S T R A C T — Post-prosthetic complications during dental implantation are accompanied by pain symptoms leading to disorders of the psychoemotional state. All this influences the behavior of patients. In addition, psychoemotional stress is often a factor of provocation and persistence of the complications. The presence of a stress state in the identified pathology, as well as the influence of various therapies on the treatment of post-prosthetic complications, is reflected in the dynamics of changes in the concentrations of both catecholamines (epinephrine, norepinephrine) and glucocorticoids (cortisol) — hormones of the medulla and the adrenal cortex. The aim: to conduct an immunological analysis of the stress response in patients with post-prosthetic complications during dental implantation. Materials and methods: The study was performed in 120 patients with post-prosthetic complications during dental implantation before and during treatment: Group I (control) — 30 patients treated with the conventional therapy; group II — 30 patients treated with ozone therapy in addition to the conventional therapy; group III — 30 patients treated with transcranial electrical stimulation in addition to the conventional therapy; Group IV — 30 patients treated with a combination of conventional therapy, ozone therapy and transcranial electrical stimulation. The concentration of epinephrine, norepinephrine, cortisol, alkaline phosphatase, and the Garkavi index were evaluated. Results: The change in these indicators after the treatment indicates the normalization of the level of the studied enzymes — markers of bone homeostasis, which is confirmed by an improvement in the clinical picture in the oral cavity. Conclusion: changes in immunological parameters objectively reflect the psychoemotional state of patients. The nature of changes in the hormonal stress response to the treatment of post-prosthetic complications indicates the effectiveness of the therapeutic regimens used, and, as a result, a decrease in both pain symptoms and psychoemotional stress.

K E Y W O R D S — epinephrine, norepinephrine, inflammation, implants, treatment.

Yuliya Makedonova1,2 , Dmitry Mihalchenko1 ,Oksana Kurkina1 , Olga Afanasyeva1 ,Sergey Veremeenko1 , Elena Venskel1

1 Volgograd State Medical University, Volgograd2 Volgograd Medical Research Center, Volgograd, Russia

[email protected]

Received 11 March 2021; Received in revised form 25 April 2021; Accepted 3 May 2021

R E L E V A N C EAs a response to the impact of such aggravating

factors as pain, blood loss [7], mechanical tissue dam-age [1–5], hypoxia [6], increased free radical oxida-tion [8, 9], a non-specific neuroendocrine reaction appears [10]. Consequently, the influence of different types of therapeutic approaches for the treatment of post-prosthetic complications is reflected in the dynamics of changes in serum concentrations of both catecholamines (epinephrine, norepinephrine) [11] and glucocorticoids (cortisol) — hormones of the medulla and the adrenal cortex [12]. Determining the level of cortisol is an important indicator in the diag-nosis of a stressful situation. Increased cortisol levels are observed with a decrease in psychomotor activity, sleep disorders, and depression. The reference values of cortisol in the blood serum are 200–700 nmol/l at 8.00 a.m. and 55–250 nmol/l at 20.00 pm. The differ-ence between the concentrations in the morning and in the evening should be at least 100 nmol/l.

Тhe biochemical catalyst for stress is epinephrine, which is released by the adrenal glands. It activates gly-cogenolysis in muscle cells, rises heart rate and blood pressure, dilates blood vessels of heart muscles and stimulates the production of ACTH and glucocorti-coids. The level of adrenaline in the blood depends on the tone of the sympathetic system. Its content in the blood plasma should not exceed 112–658 pg/ml.

The mediator of the sympathetic nervous system is the hormone of the adrenal medulla — nore-pinephrine. A decrease in the level of the ratio of norepinephrine to adrenaline in the urine of patients may indicate their depressive states. In severe de-pression, the level of the norepinephrine metabolic product - 3-methoxy-4hydroxyphenylglycol is often reduced, and the amount of the serotonin deriva-tive — 5-hydroxyindolacetic acid is reduced in the cerebrospinal fluid of these patients. An increase in the daily excretion of epinephrine and a decrease in the release of norepinephrine indicate a depressive phase of manic-depressive psychosis, and a manic state is characterized by a multiple increase in the excretion of norepinephrine. Both in mania and in depression, the blood content of adrenaline in patients is increased.

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/24

https://orcid.org/0000-0002-5546-8570

https://orcid.org/0000-0002-0479-8588

https://orcid.org/0000-0002-4759-3777

https://orcid.org/0000-0001-8577-2939

https://orcid.org/0000-0003-0652-0173

https://orcid.org/0000-0002-4479-9729

http://dx.doi.org/10.35630/2199-885X/2021/11/2/24


A decrease in the level of norepinephrine leads to an increase in the level of cortisol, and a deficiency of serotonin, which is accompanied by a disruption in regulation of cortisol secretion. The lack of both leads to disruption of circadian rhythms in the secretion of cortisol. The reference values of the content of norepinephrine in the blood plasma should not exceed 10 pg/ml.

The aim of our study was to evaluate the stress response of patients with inflammatory postprothetic complications against the background of various pharmacotherapy methods according to immunologi-cal parameters.

M A T E R I A L S A N D M E T H O D SThe study included the diagnosis and follow-

up of 120 patients aged 18–44 years (WHO aver-age age) with post-prosthetic complications during dental implantation before and during treatment. The patients were randomized into groups according to the treatment regimen: Group I (control) — 30 patients who were treated with conventional treatment; group II — 30 patients who were treated with ozone therapy in addition to conventional treatment.; Group III — 30 patients, in the complex treatment of which, in addition to traditional therapy, transcranial electros-timulation was performed; group IV — 30 patients, whose complex treatment included a combination of traditional therapy, ozone therapy and transcranial electrostimulation.

Based on the general and biochemical blood analysis, the indicators that allow us to assess the stress-ful situation in the study groups (cortisol concentra-tion, alkaline phosphatase, Garkavi index, epinephrine, norepinephrine) were determined.

General and biochemical blood tests were per-formed using a laboratory test. The leukocyte formula, the concentration of glucose, creatinine, total protein and other indicators were studied. The type of adaptive response (stress, quiet activation, re-activation) was determined by the percentage of lymphocytes in the peripheral blood, taking into account age. The Garkavi index was used as a basis.

Garkavi index = lymphocytes (%) / segmental neutrophils (%);

In healthy adults, this indicator is within the values of 0.3–0.5.

To determine the alkaline phosphatase (ALP) in the blood serum, a spectrophotometric method was used using reagents from the company "Human" (Germany) and a calibration curve constructed from a standard solution of p-nitrophenol. The method is based on the following reaction:

alkaline phosphatase

p-nitrophenyl phosphate + H2O – " p-nitrophe-nol+ phosphate

The measurement was carried out at a tem-perature of 37° C and a wavelength of Hg 405 nm (400–420 nm). The average change in optical density per minute (AA/min) was calculated. The activity of the alkaline phosphatase was calculated by the formula

E / l = AA/min x 2757Blood samples of the patients in the volume of 5

ml were obtained from 6.00 to 10.00 a.m. on an empty stomach (Fig. 1).

On the eve of the examination, patients were advised to refrain from overeating, eating salty, spicy, fatty foods, exclude alcohol, and do not smoke an hour before the examination. The biomaterial was collected before undergoing instrumental examination and physiotherapy. Venous blood obtained without anticoagulants in a centrifuge glass tube is settled in it at room temperature, then centrifuged for 10 minutes at 2500 rpm.

Repeated blood tests to monitor the dynamics of indicators were carried out under the same conditions (time, food regime) and in the same laboratory, since the blood sampling algorithm, study methodology and reference values (norms) may differ significantly in different medical institutions.

The results were evaluated on day 3, 7, 14, and 1 month later. Two-factor analysis of variance (RANOVA) with the Newman-Keuls post-hawk test was performed; * — p<0.05; ** — p<0.01;*** — p<0.001.

The clinical trial was approved by the Regional Ethics Committee, Protocol No. 2115/1-2019 of April 19, 2019.

R E S U L T SBefore the start of therapy, no statistically

significant differences in the studied parameters were found between the groups. At the same time, for all certain hormones, a statistically significant decrease was observed as a result of therapy. The study revealed a relative increase in the concentrations of hormones (epinephrine, norepinephrine, cortisol) before the start of therapy, which reflects the overall clinical picture of the stress response. At the same time, there were no statistically significant differences between the groups before the start of therapy. The content of epine-phrine varied in the range of 127.047±19.4024 pg/ml, norepinephrine 381.3607±40.2137 pg/ml, cortisol 430.62±40.275 nmol/L.

When determining the content of epinephrine in all groups, there was a decrease in its concentration at a more intense rate following the combined method of treatment (Fig. 2).

D E N T I S T R Y


Fig. 1. Venous blood sampling

Fig. 2. Dynamics of changes in the concentration of epinephrine during 1 month of observation

The decrease in the concentration of epinephrine in the blood serum occurred up to the 5th control point (1 month). At the same time, the treatment regi-men combined with ozone therapy showed a signifi-cantly greater decrease (59.348 vs 73.438; p<0.01), but for groups III and IV, the differences were even higher (59.348 vs 58.348 and 55.406, respectively, p<0.001). At the same time, in the nearest follow-up period (up to 14 days), the lowest values were typical for groups II and IV (p<0.01 and p<0.001, respectively). For norepinephrine, similar dynamics were observed, while the differences with the comparison group (conventional therapy) were less pronounced. At the same time, the dynamics of the greatest decrease in the concentration of norepinephrine in the immediate follow-up period (3–30 days) was shown for the treat-ment regimen combined with ozone therapy (3, 14, 30 days — p<0.01; 7 days — p<0.001), similar values were found for group IV (3, 7, 14, 30 days — p<0.01). The dynamics of changes in concentrations characteristic of epinephrine and norepinephrine and the revealed tendency to decrease in the near-term follow-up were also shown for changes in serum cortisol concentra-tions, regardless of therapeutic regimens. And by day 14 of the therapy they were significantly lower relative to the start of therapy and 3 days (p<0.05). This was followed by a slight increase in these parameters. The inter-group differences for cortisol were similar to those for norepinephrine, but the lowest values were observed at the 4th control point for all groups. The dynamics of the greatest decrease in cortisol concen-tration in the nearest follow-up period (3–30 days) was observed in group IV (7, 14, 30 days — p<0.01; 3 days — p<0.001). Similar values were found for group II (3, 7, 14, 30 days — p<0.01). An increase in the Garkavi index which reflects the relationship between the humoral and cellular components of immunity, an assessment of the stress state, as well as an assessment of the adaptive responses of training and activation, was found in all study groups, whereas no significant differences were found between the groups. The aver-age value was 0.687±0.05. In the course of all types of therapy normalization of the index was achieved by 14 days. A stable decrease in the index in all groups during the first 2 weeks indicates a decrease in the active in-flammatory process. Thus, the nature of changes in the hormonal stress response to the treatment of post-pros-thetic complications indicates the effectiveness of the therapeutic regimens used, and, as a result, a decrease in both pain symptoms and psychoemotional stress. In the near term of observation, there is a significant decrease in the concentrations of the studied hormones (catecholamines and corticosteroids), normalization of the values of the Garkavi index (Table 1–4).

The concentration of epinephrine in group I significantly decreased until 1 month after the start of therapy.

Evaluating the Garkavi index, a stable decrease in the index was revealed in all groups for 2 weeks which indicates a decrease in the active inflammatory process. Normal values of the index were maintained on day 30.

The parameters of epinephrine concentration in group II (in the complex treatment, ozone therapy was additionally used), group III (additional use of transcranial electrical stimulation) and in group IV (a combination of methods of groups 1–3) behaved similarly to the first, but the decrease to 1 month from the start of therapy was more pronounced.

Thus, the analysis of the dynamics of the content of stress hormones in the blood serum showed a char-acteristic and significant decrease in the concentra-tions of hormones up to 1 month, followed by a slight increase in concentrations. These trends indicate the effectiveness of the therapeutic regimens used, and as a result, a decrease in both pain symptoms and psychoe-motional stress. And taking into account the relatively

D E N T I S T R Y


maximum decrease in the concentrations of the pre-sented hormones for group IV, combining traditional therapy with ozone therapy and transcranial electrical

stimulation, for which there were large inter-group differences relative to traditional therapy both in the near and long-term follow-up periods, it can be argued

Signs/deadlines

Cortisol SCHF Index Garkovi Adrenaline rush NorepinephrineM SD M SD M SD M SD M SD

Before treatment 420,400000 38,760000 118,00000 10,75200 0,687400 0,05000 127,35840 20,14200 381,348600 40,354800

3 days 362,286300 30,698040 122,59390 8,75520 0,561200 0,03470 105,34580 16,35400 362,286300 20,2528207 days 266,626500 26,680250 122,56790 6,32016 0,523040 0,03280 94,86240 16,43800 266,626500 22,80148014 days 215,480000 19,364800 133,25590 9,21600 0,437300 0,03760 86,96423 15,34600 215,480000 18,2587901 month 29,881800 20,482120 145,99580 9,02880 0,450300 0,05380 73,43840 13,24580 219,881800 17,504430

Signs/deadlines

Cortisol SCHF Ind.Garkovi Adrenaline rush NorepinephrineM SD M SD M SD M SD M SD



Signs/deadlines




Signs/deadlines




Table 1. Immunological examination of blood in patients of group I

Table 2. Immunological examination of blood in patients of group II

Table 3. Immunological examination of blood in patients of group III

Table 4. Immunological examination of blood in patients of group IV

D E N T I S T R Y


that this scheme was most effective in the treatment of post-prosthetic complications.

An increase in the content of alkaline phos-phatase up to 1 month and a further retention of the values in the plateau phase may indicate that the use of therapy, especially combined with ozone therapy and transcranial electrical stimulation, activates the proc-esses of connective tissue synthesis.

The results of the obtained values indicate a high level of psychological stress in groups of patients with different therapy for post-prosthetic complications of dental implantation at different time intervals. Based on the data obtained, it can be argued that the changes in the concentrations of epinephrine, norepinephrine, alkaline phosphatase, and cortisol detected at the beginning of therapy fully fit into the picture of the immune response under psychophysiological stress and objectively reflect the changes in the psychophysi-ological state of patients with postprothetic complica-tions.

R E F E R E N C E S1. Avanisyan V., Al-Harazi G., Kondratyeva

T., Harutyunyan Yu. Morphology of facial skeleton in children with undifferentiated connec-tive tissue dysplasia. Archiv EuroMedica. 2020. Vol. 10; 3: 130–141. https://dx.doi.org/10.35630/2199-885X/2020/10/3.32

2. Davydov B.N. Peculiarities of microcirculation in periodont tissues in children of key age groups suffi-cient type 1 diabetes. Part I. Periodontology, 2019; Vol. 24; 1–24(90): 4–10. DOI: 10.25636/PMP.1.2019.1.1

3. Davydov B.N. Peculiarities of microcirculation in periodont tissues in children of key age groups sufficient type 1 diabetes. Part II. Periodontology. 2019;24(2):108–119. (In Russ.) DOI:10.33925/1683-3759-2019-24-2-108-119

4. Harutyunyan Yu. Undifferentiated connective tissue dysplasia as a key factor in pathogenesis of maxil-lofacial disorders in children and adolescents. Archiv EuroMedica. 2020. Vol. 10; 2: 83–94. https://dx.doi.org/10.35630/2199-885X/2020/10/2.24

5. Davydov B.N. Clinical and functional approaches to comprehensive treatment of periodontal dis-eases in children with type I diabetes. Parodon-tologiya. 2021;26(1):9–19. (In Russ.) https://doi.org/10.33925/1683-3759-2021-26-1-9-19

6. Khan M.A., Krestyashin V.M., Vakhova E.L., Krestyashin I.V., Zimina O.Yu. Тhe use of physical therapy for knee injuries in children. Archiv EuroMedica. 2019. Vol. 9. № 1. Р. 116–118.

7. Makedonova Yu. A., Mikhalchenko D. V., Dy-achenko D. Yu., Veremeenko S. A. Hemomicro-circulation of tissues around dental implants: clinical and functional parallels // Periodontology. 2020. Vol. 25. no. 4. pp. 338–342. DOI: 10.33925/1683-3759-2020-25-4-338-342.

8. Makedonova Yu. A., Mikhalchenko D. V., Zhidovinov A. V., Dyachenko D. Yu., Ver-emeenko S. A. Comparative evaluation of treat-ment efficiency of inflammatory complications after orthopedic treatment with up-to-date methods of pharmacotherapy // Journal of International Dental and Medical research. – 2020. – 13(2) – 571–576.

9. Basov A.A., Ivchenko L.G., Nuzhnaya C.V. The role of oxidative stress in the pathogenesis of vascular complications in children with insulinable sugar dia-betes // Archiv EuroMedica. 2019. Vol. 9; 1: 136–145. https://doi.org/10.35630/2199-885X/2019/9/1/136

10. Mikhalchenko D.V., Vorobyev A.A., Alex-androv A.V., Makedonova Yu.A., Shkarin V.V. Мicrohemodynamic changes as indicator of psychoеmotional stress at dental treatment. Archiv EuroMedica. 2020. Vol. 10. No 2. Р. 102–105.

11. Mikhalchenko D.V., Makedonova Yu.A., Adamovich E.I., Marymova E.B., Venskel E.V. LDF-metry as a method of diagnosing the psycho-physiological state of the body at a dental appoint-ment. Clinical dentistry. – 2020; 3 (95): 66–71. DOI: 10.37988 / 1811-153X_2020_3_66

12. Sadykov M.I., Nesterov A.M., Domenyuk D.A., Ertesyan A.R., Konnov V.V., Matrosov V. Biomechanical evaluation of stress -strain condition of restorative ceramic pin structures and dental roots. Archiv EuroMedica. 2020. Vol. 10. № 2. Р. 115–120. https://doi.org/10.35630/2199-885X/2020/10/2.29

D E N T I S T R Y


DEVELOPMENT OF A BIODEGRADABLE MATERIAL FOR BONE REGENERATION BASED ON SILICON COMPOUNDS

A B S T R A C T — In the course of this study we established that the laboratory sample in granulated form developed by us has a stimulating effect on the growth and proliferation of osteoblasts. Analysis of the data (CT, MRI) of the biological material obtained from the animals withdrawn from the experiment showed a statistically significant replacement of the artificial defect with bone tissue with signs of restoration of the cortical plate. The analysis of the CT results indicates that this method is sufficiently informative in assessing the regeneration of the trabecular structure of bone tissue. The study (MRI) of the bone material obtained from the animals participating in the experiment showed that when the osteoinductive material was integrated, the defect zone was replaced by 90%. In the projection of the defect, the newly formed tissue of a homogeneous structure was determined, corresponding in density to bone tissue with a high degree of mineralization. There was a complete restoration of bone tissue in the projection of the defect.

K E Y W O R D S — osteoplastic materials, bone defects, bone powder, dental implantation.

Dmitry Kompantsev , Anna Chahirova ,Ruslan Yusupov, Ilya Zaharchanko,Natalia Shabanova

Pyatigorsk Medical and Pharmaceutical Institute —branch of Volgograd State Medical University, Pyatigorsk, Russia

[email protected]


donor sites (ilium, ribs, mandible), complication and lengthening of surgical intervention, increased risk of complications, the specifics of the somatic status and preferences of patients impose restrictions on the use of autogenous bone tissue and force the use of prod-ucts available on the market.

However, the vast majority of osteoplastic materials on the market are a kind of biologically active additives — they only supply structural components necessary for the formation of bone tissue in the defect zone, therefore they have low or extremely moderate effectiveness.

Purpose of the studyThe purpose of this study was to establish the

regularities of morphological changes in bone tissue of laboratory animals for correction of defects in the maxillofacial region using osteoplastic material.

M A T E R I A L S A N D M E T H O D SExamination (CT, MRI) of biological material

obtained from animals were used in the experiment. The results of the experiment were registered.

In our experiments we fully adhered to Ethical Guidelines for the Use of Animals in Research (EU Directive 2010/63/EU).

The object of the CT study was the bone tissue of the lower jaw of a rabbit obtained 90 days after the integration of experimental osteoplastic material, fixed in 10% buffered formalin in accordance with the rules for pathomorphological and histological studies.

There was a complete restoration of bone tissue in the projection of the defect (Fig. 1). The newly formed bone looked immature: the bone beams were uneven in shape and size, randomly arranged.

In CT with the reconstruction of the three-dimensional image, the replacement of the defect with bone tissue with the restoration of the cortical plate was determined. The CT data allowed us to visualize the formation of the trabecular structure of bone tissue in the projection of the defect at a given regeneration period of 90 days (Fig. 2).

The analysis of CT results in bone regeneration showed its informative value in assessing the trabecular

I N T R O D U C T I O NOne of the most urgent problems of traumatol-

ogy, orthopedics, surgical dentistry and maxillofacial surgery is the problem of replacing bone defects. A distinctive feature of these problems is the lack of expression of the natural regenerative processes of bone tissue. Given that the spontaneous restoration of the integrity of bone tissue in the area of the defect is either impossible or excessively long, therapeutic meas-ures are based on the use of various bone-substituting (osteoplastic) materials, which are designed to enhance the activity and effectiveness of the recovery process.

The most effective of the osteoplastic materials is the patient's own (autogenous) bone tissue. However, the need to expand or create a new surgical field to obtain fragments of the patient's own bone tissue from

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/25

https://orcid.org/0000-0002-5074-808X

https://orcid.org/0000-0002-5265-9478

https://orcid.org/0000-0002-7693-5182

http://dx.doi.org/10.35630/2199-885X/2021/11/2/25


Fig. 1. Three-dimensional 3D model of bone tissue in color

Fig. 2. Formation of the trabecular structure of bone tissue. Three-dimen-sional 3D model in color

structure of bone tissue. The reconstruction of three-dimensional images made it possible to determine the spatial localization of structures in the projection of the bone defect due to the absence of the overlap effect, to outline the surface structures in relief and to evaluate the complex three-dimensional relationships between bone and soft tissue fragments.

The study (MRI) of the biological material ob-tained from the animals in the experiment showed that when the osteoinductive material was integrated, the defect zone was replaced by 90%.

In the projection of the defect, the newly formed tissue of a homogeneous structure was determined, corresponding in density to bone tissue with a high degree of mineralization. There was a complete res-toration of bone tissue in the projection of the defect (Fig. 3). The newly formed bone looked immature: the bone beams were uneven in shape and size, randomly arranged.

Histological examination of the bone in the defect area of the lower jaw of rabbits in the time interval from 14 days till 3 months has shown that the healing of the bone wound, the features and the rate of formation and maturation of bone regenerate were as-

Fig. 3. MRI, three-dimensional image reconstruction

Fig. 4. 3D volumetric model in color

sociated with the osteoinductive and osteoconductive properties of the experimental drug.

C O N C L U S I O NControl studies of the biological material

obtained from the animals used in the experiment confirmed our research.

The results of computed tomography revealed the persistence of the tendency to increase the density of regenerate in animals. By the end of the experiment, the bone defect was completely closed. Along the edges of the former opening, the trabeculae were par-tially resorbed and the bone began to transform into a lamellar bone with a developed system of haversoid channels. Bone tissue rearrangement in the area of the formed regenerate in animals with local administration of the experimental drug occurred at an earlier time, which was determined by the density of the regenerate, this is a sign of a sufficiently high degree of biocompat-ibility of bone tissue and experimental osteoplastic material (Fig. 4).

The MRI results confirmed that the implanted material corresponds to the bone tissue in density and structure, and the regenerate density is uniform (Fig. 5).

D E N T I S T R Y


On the 14th day after the operation, the forma-tion of immature bone tissue began, by the 20th day, the bone tissue already filled most of the volume of the defect (two-thirds). At the same time, there was a cer-tain maturation of the regenerate, characterized by an orderly arrangement of fibrous structures, although it still consisted of osteoid beams. Part of the regenerate had a chondroid structure. 1 month after the opera-tion, the defect was almost completely filled with bone regenerate, the bone marrow was absent (Fig. 6). After 2 months, the reticulofibrous bone tissue matured and was partially replaced by lamellar bone. After 3 months, the bone tissue of the regenerate has already fully matured, compacted, clear lines of adhesion ap-peared, and osteons were formed (Fig. 7).

The bone regenerate was similar in structure to the spongy bone of the operation area, only slightly different in thickness and pronounced signs of bone remodeling.

R E F E R E N C E S1. Deev R.V., Drobyshev A.Y., Bozo I.Y., Isaev

A.A. Ordinary and activated bone grafts: ap-plied classification and the main features //Bi-oMed Research International. – 2015. – Vol. 2015. Doi:10.1155/2015/365050

2. Aslan M., Şimşek G., Dayi E. The effect of hyaluronic acid-supplemented bone graft in bone healing: experimental study in rabbits //Journal of biomaterials applications. – 2006. – Vol. 20. – No. 3. – P. 209–220. Doi: 10.1177/0885328206051047

3. Dahiya P., Kamal R. Hyaluronic acid: a boon in periodontal therapy //North American journal of medical sciences. – 2013. – Vol. 5. – No. 5. – P. 309. Doi: 10.4103/1947-2714.112473.

4. Schulz M. C., Korn P., Stadlinger B., et al. Coating with artificial matrices from collagen and sulfated hyaluronan influences the osseointegration of dental implants //Journal of Materials Science: Materials in Medicine. – 2014. – Vol. 25. – No. 1. – P. 247–258. Doi: 10.1007/s10856-013-5066-3.

Fig. 5. MRI of the bone tissue of the lower jaw

Fig. 6. The defect is filled with maturing regenerate, the bone compaction looks weak. Stained with hematoxylin and eosin

Fig. 7. Relatively mature bone regenerate with compaction phenomena. Stained with hematoxylin and eosin

5. Chang YL, Lo YJ, Feng SW, et al. Bone healing improvements using hyaluronic acid and hydroxya-patite/beta-tricalcium phosphate in combination: an animal study //BioMed research international. – 2016. – Vol. 2016. Doi:10.1155/2016/8301624

D E N T I S T R Y


THE PREVALENCE OF NON-CARIOUS TOOTH DEFECTS ASSOCIATED WITH INDUSTRIAL EXPOSURE IN RESIDENTS OF PENZA REGION (RUSSIA)

A B S T R A C T — Aim of the study: to assess the level of dental pathology in Penza Region (Russia) and estimate the impact of the industrial environment. Materials and methods: A clinical examination and analysis of the frequency of enamel erosion, fluorosis and hypoplasia, wedge-shaped defect and multiple enamel cracks were carried out among the main professional groups of the region's population — workers of the machine-building and timber processing industries in the Penza region. Results and Discussion. Among the residents of Penza region, the frequency of enamel erosion was 4.0%, pathological tooth wear — 7.1%, wedge-shaped defect — 6.1%, multiple enamel fractures — 11.1%. There is a high frequency of non-carious tooth defects that occur before teething. In patients working in the machine-building industry, a high frequency of pathological tooth wear was noted (24,5%); in workers of the timber processing industry, it was 2 times lower (12,6%), as well as a high frequency of wedge- shaped defects, enamel erosion and multiple enamel cracks — all this was 2–2.5 times higher compared to similar indicators in patients from agricultural populations. However, the prevalence of dental fluorosis and hypoplasia was almost the same as in the agricultural population. Conclusion: Penza region has a high prevalence of dental fluorosis and hypoplasia. At the same time, the influence of industrial environment on the incidence of various types of non-carious dental pathology in workers of machine-building and timber processing industries in Penza region was established.

K E Y W O R D S — non-carious tooth defects (NCTD); prevalence; intensity; concentration of fluoride.

Marina Suvorova , Galina Emelina ,Anna Teplova , Belousova Anna

Penza State University, Penza, Russia

[email protected]


mal enamel and its preservation [6]. The differential diagnosis of enamel abnormalities should be carried out with various forms of dental fluorosis, which also manifests itself as a variety of white, yellowish or brown spots on several or all teeth [5]. Fluorosis is an endemic disease that occurs as a result of an increased content of fluoride in drinking water. The exact mechanism of the occurrence of fluorosis is not yet fully understood. It is believed that fluorosis increases the content of fluoride in saliva and on the surface of enamel. It was shown that the prevalence of fluorosis directly depends on the content of fluoride in drink-ing water: at a fluoride concentration of 0.8–1.0 mg/l, the prevalence of fluorosis is 10–12%, 1.0–1.5 mg/l — 20-40%, 1.5–2.5 mg/l — 30–40%, over 2.5 mg/l — more than 50% [2]. Thus, the intensity of fluorosis in the population in endemic foci can be different: from mild lesions to significant changes in the teeth. This means that at the same concentration of fluoride in water, the body can react differently to its intake [1].

M E T H O D SA clinical examination and analysis of the fre-

quency of enamel erosion, fluorosis and hypoplasia, wedge-shaped defect and multiple enamel cracks were carried out among the main professional groups of the region's population-employees of the machine-building and timber processing industries of the Penza region.

In this study, we analyzed dental examination data from 200 subjects aged 25 to 75 years (56 men and 144 women), of which 16 people were aged 21 to 30 years, 40 people — 31 to 40 years, 24 people — 41 to 50 years, 80 people — 51 to 60 years, 24 people — 61 to 70 years, 16 people — 71 to 80 years.

R E S U L T S Among the native residents of Penza region (rep-

resented by the group of the rural population almost completely, without the share of the alien population), the frequency of enamel erosion was 4.0%, pathologi-cal tooth wear was 7.1%, and the wedge-shaped defect was 6.1%. Occurrence of multiple fractures of the enamel was even more common — 11.1%. Attention is drawn to the high frequency of non-carious dental diseases that occur before eruption (the incidence of

I N T R O D U C T I O NIn addition to caries, a significant place among

dental diseases is occupied by non-carious enamel lesions: various types of hypoplasia (systemic, local, fo-cal), imperfect amelogenesis, dentinogenesis, Stanton-Candepon syndrome, as well as fluorosis.

Non-carious lesions of the enamel of the teeth, which have a congenital character, are formed in the antenatal period. The physiological course of preg-nancy is of great importance in the biosynthesis of nor-

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/26

https://orcid.org/0000-0001-5601-2869

https://orcid.org/0000-0002-6085-5591

https://orcid.org/0000-0002-5785-5515

https://orcid.org/0000-0001-8223-2697

http://dx.doi.org/10.35630/2199-885X/2021/11/2/26


dental hypoplasia was 14.1%, dental fluorosis 24.2%) — the region is endemic for fluorosis.

Interesting differences in the incidence of major non-carious tooth defects are provided by an analysis of respondents from the major occupational groups of the population – workers of machine-building and woodworking industries. In patients from the machine-building industry, there was a high incidence of pathological tooth wear (24.5%, and in workers of the timber processing industry — 12.6%, 2 times lower), as well as a high incidence of wedge–shaped defects (13.3%), enamel erosion (11.4%) and multiple enamel cracks (20.9%) — all this was 2–2.5 times higher compared to similar indicators in patients from the agricultural population.

However, the prevalence of fluorosis and dental hypoplasia was almost the same as in the agricultural population — 13.3% and 11.4% (even the incidence of fluorosis was slightly lower, which indicates a greater proportion of the non-indigenous population of the region in the group of workers in the engineer-ing industry). In patients from workers of the timber processing industry, a high frequency of many non-carious diseases of the teeth was also noted, but less than in workers of the machine-building industry: in particular, the frequency of occurrence of pathological tooth wear was 12.6%, wedge-shaped defect — 8.0%, enamel erosion — 8.0%, multiple enamel cracks — 14.9%. High prevalence rates of fluorosis (13.8%) and dental hypoplasia (8.0%) were observed.

Overall, it was noted that on the first place in frequency of non-carious tooth defects among the workers of machine-building industry was pathologi-cal tooth wear (24,5%) followed by multiple fractures of the enamel (20,9%) and wedge-shaped defect and fluorosis of the teeth (13.3%), and more rarely — the erosion of enamel hypoplasia of the teeth (11.4%). Among the workers of the timber processing industry, multiple enamel cracks (14.9%) and dental fluoro-sis (13.8%) were most often encountered, less often — pathological tooth wear (12.6%), wedge-shaped defect, enamel erosion, tooth hypoplasia (8.0% each). In respondents employed in agriculture, non-carious pathology was mostly manifested as fluorosis (in 24.2% of respondents in this group) and hypoplasia of teeth (14.1%). Multiple enamel cracks (11.1%), pathological tooth abrasion (7.1%), wedge-shaped defect (6.1%) and enamel erosion (4.0%) were less common.

D I S C U S S I O NIn other words, work in the machine-building

industry of the Penza region features the highest inci-dence frequency of certain types of non-carious tooth defects. In this population pathological tooth wear was

2 times higher than in workers of the timber process-ing industry and almost 4 times higher than in people employed in agriculture. The wedge-shaped defects and enamel erosion was also 1.5 times higher com-pared with workers of the timber processing industry and 2 times higher than in respondents employed in agriculture. On the other hand, agricultural popula-tions had a higher incidence of dental fluorosis and hypoplasia, which is associated with a higher propor-tion of the native population and a smaller propor-tion of the alien population. The leading role in the frequency of different types of non-carious pathology among workers of the machine-building and wood-processing industries is occupied by pathological tooth wear, wedge-shaped defect and multiple enamel cracks, which proves the influence of industrial hazards.

Analysis of correlation between the frequency of non-carious tooth defects and the seniority of different groups of workers allowed us to determine the depend-ence of this disease from exposure to occupational factors. Thus, high indicators of strong correlation relationship were noted in the groups of workers from machine-building and timber processing industries in relation to the pathological tooth wear (0.76 and 0.70 points) and the wedge-shaped defect (0.68 and 0.64 points); these indicators were almost 2 times lower for those employed in agriculture (0.37 and 0.30 points). In addition, high values of the correlation with senior-ity were demonstrated in respondents employed in the machine-building and timber processing indus-tries, specifically in regard to enamel erosion (0.61 and 0.55 points) and multiple enamel cracks (0.79 and 0.76 points). Among agriculture workers, signifi-cantly lower values were noted (0.18 and 0.16 points, respectively). It should be noted that such disorders as fluorosis and dental hypoplasia did not show the influence of seniority on the frequency of occurrence: the correlation coefficients in respondents employed in the machine-building and timber processing industries were 0.16 points and 0.17 points, respectively; 0.15 points and 0.11 points. For almost all non-carious tooth defects, low correlation coefficients of work ex-perience were observed in the agricultural population.

C O N C L U S I O NThus, the analysis of the regional features in the

incidence of non-carious tooth defects in the residents of the Penza region showed the following. The region is endemic for fluorosis, which is why there are high prevalence rates of fluorosis and hypoplasia of teeth. At the same time, the influence of industrial environment on the incidence of various types of non-carious tooth defects in the workers of machine-building and timber processing industries in the Penza region was established.

D E N T I S T R Y


R E F E R E N C E S1. Atalay C., Ozgunaltay G., Yazici A.R. Thirty-

six-month clinical evaluation of different adhesive strategies of a universal adhesive. Clin Oral Investig. 2020 Apr;24(4):1569–1578. doi: 10.1007/s00784-019-03052-2.

2. Badavannavar A.N., Ajari S., Nayak K.U., Khi-jmatgar S. Abfraction: Etiopathogenesis, clinical aspect, and diagnostic-treatment modalities: A review. Indian J Dent Res. 2020 Mar-Apr; 31(2):305–311. doi: 10.4103/ijdr.IJDR_863_18.

3. Caneppele T.M., Meirelles L.C., Rocha R.S., Gonçalves L.L., Ávila D.M., Gonçalves S.E., Bresciani E. A 2-year clinical evaluation of direct and semi-direct resin composite restorations in non-carious cervical lesions: a randomized clinical study. Clin Oral Investig. 2020 Mar;24(3):1321–1331. Epub 2019 Jul 11.PMID: 31297659 Clinical Trial. doi: 10.1007/s00784-019-03011-x

4. Du J.K., Wu J.H., Chen P.H., Ho P.S., Chen K.K. Influence of cavity depth and restoration of non-car-ious cervical root lesions on strain distribution from various loading sites. BMC Oral Health. 2020 Apr 7;20 (1):98. doi: 10.1186/s12903-020-01083-w.

5. Emelina G. V., Suvorova M. N., Gerashchenko S. M., Gerasimova T. V., Emelina E. S. Compara-tive analysis of dental morbidity as the basis of popula-tion demand for dental services. Journal of Pharma-ceutical Sciences and Research. 10(4), P. 798–799.

6. Emelina G. V., Suvorova M. N., Zyulkina L. A., Kuznetsova N. K., Emelina E. S. Evaluation of the incidence of inflammatory processes in periodontal tissues in the Penza region. 2018. Journal of Pharma-ceutical Sciences and Research. 10(4), pp. 973–975. 4.

7. Lim G.E., Son S.A., Hur B., Park J.K. Evaluation of the relationship between non-caries cervical lesions and the tooth and periodontal tissue: An ex-vivo study using micro-computed tomography. PLoS One. 2020 Oct 28;15 (10):e0240979. doi: 10.1371/journal.pone.0240979.

8. Sevbitov A.V., Enina Yu. I., Dorofeev A. E., Kamenskov P. E., Kozhemov S. I., Nikonova A.V. Investigation of the influence of various abrasive factors on the surface microrelief of hybrid ceramic orthopedic structures. An option. 2019. Vol. 35. no. S24. p. 598–611.

D E N T I S T R Y


IMPLANT TREATMENT IN PATIENTS WITH SEVERE TOOTH WEAR ON THE OCCLUSAL SURFACE

A B S T R A C T — Examination along with implant treatment was carried out in 21 patients with severe tooth wear on the occlusal surface. This disorder features a variety of clinical symptoms including changes in the facial features and the lower jaw movement amplitude, disturbed canine and incisor guidance, supercontacts, asynchronous operation of the masticatory muscles, etc. We have proposed a treatment algorithm aimed at eliminating these issues.The temporary non-removable dentures allowed creating occlusal marks (Stage 1) followed with their transfer onto permanent orthopedic structure (Stage 2) using implants. The complexity of the approach to treating this disease, as well as the effectiveness of our method has been confirmed by clinical data and the outcomes obtained through an instrumental study involving electromyography and electronic axiography.The extra additional research methods employed, such as electromyography and electronic axiography, allowed to assess the activity of masticatory muscles and the lower jaw articulation, thus to ensure occlusal-articulation interaction when restoring the lower facial height.

K E Y W O R D S — increased abrasion, dentures, electromyography, electronic axiography.

Vladimir Tlustenko , Valentina Tlustenko ,Sergey Komlev , Alexander Ivaschenko ,Vladimir Koshelev

Samara State Medical University, Samara, Russia

[email protected]

Received 25 April 2021; Received in revised form 29 May 2021; Accepted 3 June 2021

function guiding the canines and incisors, whereas the patient’s own perception of the bite changes. Increased dental abrasion is an etiological factor behind the temporomandibular joint (TMJ) dysfunction [4–6], it is often accompanied by a partial adentia, which poses certain difficulties for implantation [7].

Considering the negative impact of occlusion-articulation changes due to increased dental abrasion [8], and the risk of changes in bite forces after the im-plant treatment, restoring the structure on the occlusal surface of dentures appears to play an important role [9, 10]. Lack of occlusion reference marks complicates carrying out rehabilitation measures up to their fullest extent [11–15]. In this regard, the necessary steps include proper remodeling of the occlusion, restoring the lower face height, and monitoring the function-ing of the newly designed orthopedic structures, both shortly after setting the dentures, and long-term.

Aim of the study: to improve the effectiveness of orthopedic treatment with implants in patients with increased dental abra-sion on the occlusal surface.

M A T E R I A L S A N D M E T H O D SWe examined 26 patients aged 35 to 53, among

them 16 males and 10 females, who had increased dental abrasion on the occlusal surface, complicated by partial adentia. 93 implants (Semados) were installed in the patients. The inclusion criteria for the study and further implant treatment were patients with tooth defects of Class III and IV according to Kennedy classification. The exclusion criteria were degenerative changes in the TMJ structural elements.

The data involving the respective medical records, the patients’ complaints, as well as the data obtained through clinical and instrumental examinations were analyzed. There were also examinations caried out such as the upper and lower jaw model analysis (Stratos 300 articulator), intraoral radiographs of teeth (Kodak 2000), and computed tomography imaging (Kodak 9000). The electronic axiography was per-formed with Arcus Digma II (Kavo Germany), elec-tromyography of masticatory muscles — with Synapsis (Neurotech Russia). The statistical data processing for the obtained outcomes was done on the SPSS 25

I N T R O D U C T I O NIncreased abrasion of the hard dental tissue (IDA)

on the occlusal surface entails a number of anatomical, morphological, aesthetic and functional disorders. The prevalence of the issue ranges from 6% to 14.5% [1–3], the most common signs being a decrease in the size of the crowns affecting all teeth, dentin hyperesthesia, supercontacts, alterations affecting the shape of the dental arches, as well as a shortened lower third of the face.

Significant dental abrasion on the occlusal surface involves a change in the lower jaw movement ampli-tude, which is due to a decrease in the inter-occlusal height. The patient may fail to close the teeth in the previous position; there is also a frequent loss of the

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/27

https://orcid.org/0000-0002-7429-9135

https://orcid.org/0000-0002-2756-5277

https://orcid.org/0000-0001-6491-4694

https://orcid.org/0000-0004-7645-9004

https://orcid.org/0000-0001-8479-0342

http://dx.doi.org/10.35630/2199-885X/2021/11/2/27


software package, with the arithmetic mean (M) and the mean square deviation (SD) calculated.

The differences between the samples were evaluat-ed employing the Student’s t-test as well as the Mann-Whitney-Wilcoxon U-test, while between the results in the same sample through treatment — with the Student’s paired t-test and the Wilcoxon paired test. The results were considered different at a significance value set at p<0.05.

R E S U L T S A N D D I S C U S S I O NThe patients were found to have a 8±1.24 mm

decrease in the lower face interocclusal height; deepen-ing nasolabial and chin folds; a deformed dentition; balancing supercontacts of molars in the lateral occlu-sions (right and left); canine disocclusion; disturbed incisor and canine guidance, as well as a partial adentia.

Since increased dental abrasion on the occlusal surface was accompanied by a significant decrease in the gnathic height, the restorative treatment was carried out in stages. Initially, a special orthopedic preparation was done (9) in the form of occlusal splint-mouthguards, which the patients should use for 3±0.15 months, and which allowed adjusting the TMJ elements prior to the implantation. 2 patients reported pain in the TMJ area, so they were further treated with the mouthguard, and the implantation had to be postponed.

24 patients were found to reveal no complica-tions, and they had temporary fixed dentures (Stage 1) made for them at a set occlusal height according to our manufacturing method (Invention Patent RU 2453290 C1), which ensures functions like chewing, stabilized occlusion, phonetics and aesthetics. The point of the method implies the following: temporary fixed bridge-like dentures were modeled from wax; oc-clusion analysis was performed in the Stratos 300 artic-ulator; the wax was replaced with a bis-acrylic material (Protemp 3 Garant or Luxatemp). The specific feature about these structures was that on the internal occlusal surface of the bridge-like prosthesis a fiberglass tape (Fiber Splint, Ribbond, and others) was adjusted in order to strengthen the frame of the entire structure. In case of IDA, the thickness of the chewing surface is usually sufficient enough. The manufactured struc-tures were fixed temporarily, after which the occlusion was evaluated with instrumental control performed through electronic axiography and electromyography of the masticatory muscles. The results obtained were as follows (see Table 1, Stage 1).

During the dental implantation, the temporary bridges in the implantation area were removed, with the implants installed, and then the temporary fixation was repeated for the osseointegration period. At the

end of the osseointegration, permanent dentures were installed.

In order to ensure the predictability for function-ing of the orthopedic structures aimed at long-term positive outcomes, repeated control of the occlusal balance in the arches was carried out, with the follow-ing results obtained (Table 1, Stage 2).

The values proposed by V.P. Tlustenko et. al. were accepted as the criteria of the norm [16].

At the Stage 1, the masticatory muscle electro-myographic examination testing the maximum jaw compression revealed a significant decrease in the biopotentials amplitude and an approximation to the normal values. The average amplitude indicators of the temporal muscle biopotentials were: 534.6±54.2 µV — on the right and 537.8±54.7 µV — on the left (p<0.05 compared to the norm); for the masseter muscles: 598.4±59.2 µV — on the right and 583.7±57.8 µV — on the left (p<0.01 compared to the norm). The symmetry factor (Sf ), which reflects the activity symmetry both between the two similar muscles on both sides, and between the masticatory and temporal muscles on one side, went up reaching the following values: for the like-named muscles — 0.8±0.1 (tempo-ral muscles) and 0.9±0.1 (masseter muscles, p=0.002); for the synergist muscles — 0.8±0.1 (the temporal and the masseter muscles on the right) and 0.8±0.1 (the temporal and the masseter muscles on the left), yet in all cases the value was significantly different from the norm (p<0.01). Given that, improving the occlusal relationship, the interalveolar height through temporary structures will allow arriving at significantly optimized activity of the masticatory muscles already at the preliminary treatment stage.

According to the EMG results, 1 month after the permanent prosthetics were completed (Stage 2), the indicators approached the normal values and, regarding the temporal muscles, were as follows: the average biopotential amplitudes — 523.6±52.8 µV (on the right, p=0.056 compared to the norm) and 514.9±52.9 µV (on the left, p=0.226 compared to the norm), the symmetry factor – 0.9±0.1 (p<0.001 compared to Stage 1, and p=0.002 – compared to the norm); for the masseter muscles: 551.2±54.2 µV (on the right, p=0.280 compared to the norm) and 541.1±53.9 µV (on the left, p=0.415 compared to the norm), the symmetry factor was 1.0±0.1. We believe that a slight excess in the indicators, if compared to the normal values, in certain patients, which was observed following the completion of the orthopedic treatment, could be accounted for by the remaining parafunctional activity of the masticatory muscles, which, as the modern ideas hold it, cannot be com-pletely stopped.

D E N T I S T R Y


The change in the closure of the arches caused by increased dental abrasion, has a direct link to disturbed articulation of the lower jaw, which can be diagnosed through electronic axiography (Table 2).

The graphical parameters featured a lack of symmetry, a decreased length of the trajectories (this indicative of articular head restricted mobility due to functional overloaded masticatory muscles through their parafunctions), emerging premature interdental contacts that block and change the lower jaw move-ment, disturbed coordination of the masticatory muscles and the asynchrony of their inclusion in activity. Prior to the treatment, the sagittal articular course angle exceeded the normal values, which could be due to a changed rotation-vs-progress component ratio related to opening the mouth, with the rotational component predominating.

At the temporary structure stage, the sagittal ar-ticular course angle decreased to 47.8±4.7° on the right and 48.3±4.1° on the left. The articular trajectories of the lower jaw movements appeared as smooth curves concaved downward with the following linear indica-tors: 11.7±1.8 mm — the right joint trajectory length, and 12.1±1.9 mm — the left one (p<0.001 compared to the norm). However, a number of axiograms revealed a discrepancy between the mouth-opening trajectory and the lower jaw movement trajectory at closing the mouth, whereas the discrepancy exceeded 0.5 mm.

Upon completion of orthopedic treatment, the angles of the sagittal articular course angles decreased on both sides, if compared to the initial values (p<0.001) and were 44.6±3.9° on the right and 45.4±4.3° on the left (p=0.210 and p=0.339 compared to the norm), whereas they also got to feature some relative symmetry of the right and left sides.

An examination of the Gothic angles revealed amplitude symmetry of the lower jaw interincisal point movements, both to the right and to the left. The lines

were not observed to have any shortening in the trajec-tory length, while the trajectories themselves appeared smooth. The protrusion line divided the Gothic angle into two identical parts in 85% of cases.

The obtained features pointed at synchronous performance of the masticatory muscles, as well as the absence of clinically significant dysfunction of the TMJ.

C O N C L U S I O NEmploying all the above-mentioned preparation

stages and orthopedic treatment of IDA on the oc-clusal surface, the inter-occlusal height and the central ratio of the jaws were identified optimally in view of the data obtained through electromyography and elec-tronic axiography. Individual movements of the lower jaw in three mutually perpendicular planes, as well as the lateral movement amplitudes within the physi-ologically normal value range, were arrived at. Restored trajectories of protrusion, laterotrusion, mediotrusion and lateroprotrusion further allow optimal sliding of the lower jaw articular head along the articular tuber-cle slope, whereas disturbance in these principles can affect the TMJ function.

R E F E R E N C E S1. Kazeko, L.A. Epidemiology of excessive dental attri-

tion/ L.A. Kazeko, O.A. Kruglik // Military Medi-cine. – 2010. – Issue 2 (15). P. 50–54. (in Russ.).

2. Alaraudanjoki V. Influence of Intrinsic Factors on Erosive Tooth Wear in a Large-Scale Epidemiological Study / V. Alaraudanjoki, ML Laitala, L. Tjäderhane et al. // Caries Research. – 2016. Vol. 50, No 5. – P. 508–516. DOI: 10.1159/000448292

3. Wetselaar P. The Prevalence of Tooth Wear in the Dutch Adult Population/ P Wetselaar, Jan H. Vermaire, C.M. Visscher, F Lobbezoo, A. Schuller//Caries Res. – 2016 – Vol. 50 No 6 – P. 543–550. DOI: 10.1159/000447020 DOI: 10.1159/000447020

EMG indices TdA avg. (µV)

TsA avg. (µV)

MdA avg. (µV)

MsA avg. (µV)

SfTd-Ts

SfMd-Ms

SfTd-Md

SfTs-Ms

Stage 1 534.6±54.2 537.8±54.7 598.4±59.2 583.7±57.8 0.8±0.1 0.9±0.1 0.8±0.1 0.8±0.1Stage 2 523.6±52.8 514.9±52.9 551.2±54.2 541.1±53.9 0.9±0.1 1.0±0.1 0.9±0.1 0.9±0.1Normal status 491.9±52.9 495.4±51.2 533.4±52.5 527.5±54.4 1.0±0.1 1.0±0.1 0.9±0.1 0.9±0.1p Stage 1 – normal 0.012 0.012 <0.001 0.002 <0.001 0.002 0.002 0.002p Stage 2 – normal 0.056 0.226 0.280 0.415 0.002 0.990 0.985 0.990

Table 1. EMG indices in patients at Stage 1 and Stage 2 through the orthopedic treatment, as compared with the normal values obtained through a functional test — maximum jaw compression in case of normal occlusion

Note: Td — right temporal muscle; Ts — left temporal muscle; Md — right masticatory muscle; Ms — left masticatory muscle; Sf — symmetry factor. The statistical significance is shown in terms of the deviation from the normal value through each of the stages. The difference between Stage 1 and Stage 2 in all cases — p<0.001

D E N T I S T R Y


4. Sadykov, S.B. Decompensated generalized excessive dental attrition is an etiological factor of temporoman-dibular joint dysfunctions / S.B. Sadykov, E.O. Isakov, K.M. Choybekova // Healthcare of Kyrgyzstan. — 2014. — Issue 1. — P. 96-99. (in Russ.).

5. Wegehaupt F., Gunhart N., Attin B.T. Preven-tion of erosive / abrasive enamel wear due to orange modified dietary supplements // Oral Dis. – 2011. – Vol. 17, No 5. – P. 508-514. https://doi.org/10.1111/j.1601-0825.2011.01797.x

6. Wetselaar, P. The tooth wear evaluation system: a modular clinical guideline for the diagnosis and man-agement planning of worn dentitions / P. Wetselaar, F Lobbezoo, // J. Oral. Rehabil. – 2016. – Vol. 43(1). – P. 69–80. DOI: 10.1111/joor.12340

7. Yeshiev, Abdyrakman Moldalievich, Paky-rov, Zhenishbek Karakozuevich. Attrition of hard dental tissues and methods of prevention and treatment. Science, education and culture. 2018; 6 (30): 48–50. (in Russ.).

8. Abolmasov, N.N., Prygunov, K.A., Abolmasov, N.G., Adayeva, I.A. Assessment of the occlusal-articulatory interaction of the dentition to identify etiological factors and symptoms of various pathologi-cal conditions (Part II) // Institute of Dentistry. – 2018. – Issue 2 (79). – P. 96–99. (in Russ.).

9. Zholudev, S.E. The role of special preparation of the oral cavity for dental prosthetics in persons with excessive attrition of hard dental tissues // Scientific Review. Medical sciences. – 2015. – Issue 1. – P. 174–174. (in Russ.).

10. Stenhagen K.R.,Hove L.H., Holme B., Tveit A.B. The effect of daily fluoride mouth rinsing on enamel erosive abrasive Caries wear in situ // Car-ies Res. – 2013. –Vol. 17, 3. – P. 815–822. DOI: 10.1159/000342619

11. Dmitrienko S.V. Enhancement of research method for spatial location of temporomandibular elements and maxillary and mandibular medial incisors. Archiv EuroMedica. 2019. Vol. 9. № 1. P. 38–44. https://doi.org/10.35630/2199-885X/2019/9/1/38

IndicatorSagittal articular course angle (°) Articular trajectory (mm)right left right left

Stage 1 47.8±4.7 48.3±4.1 11.7±1.8 12.1±1.9Stage 2 44.6±3.9 45.4±4.3 12.6±2.0 12.7±2.8Normal value 43.2±3.3 44.1±4.5 14.8±2.1 13.9±2.0p Stage 1 – norm 0.001 0.003 <0.001 0.004p Stage 2 – norm 0.210 0.339 0.001 0.112

Table 2. Angular and linear indicators of electronic axiography during the open-close- mouth test in patients at Stage 1 and Stage 2 of orthopedic treat-ment, if matched against normal values

Note: The statistical significance is shown in terms of the deviation from the normal value through each of the stages. The difference between Stage 1 and Stage 2 in all cases — p<0.001

12. Dmitrienko T.D. Connection between clinical and radiological torque of medial incisor at physi-ological occlusion. Archiv EuroMedica. 2019. Vol. 9. № 1. P. 29–37. https://doi.org/10.35630/2199-885X/2019/9/1/29

13. Shkarin V.V., Kochkonyan T.S., Ghamdan Al.H. Occlusal plane orientation in patients with dentofacial anomalies based on morphometric cranio-facial measurements. Archiv EuroMedica. 2021. Vol. 11; 1: 116–121. https://doi.org/10.35630/2199-885X/2021/11/1.26

14. Ivanyuta O.P., Al-Harasi G., Kuleshov D.A. Modification of the dental arch shape using graphic reproduction method and its clinical effectiveness in patients with occlusion anomalies. Archiv Eu-roMedica. 2020. Vol. 10; 4: – 190. https://dx.doi.org/10.35630/2199-885X/2020/10/4.42

15. Fischev S.B., Puzdyryova M.N. Morphological features of dentofacial area in peoples with dental arch issues combined with occlusion anomalies. Archiv EuroMedica. 2019. Vol. 9; 1: 162–163. https://doi.org/10.35630/2199-885X/2019/9/1/162

16. Tlustenko, V.P., Sadykov, M.I., Trunin, D.A., Potapov, V.P., Nesterov, A.M., Golovina, E.S.: Electromyography of chewing muscles: monograph. Samara: As Gard Publishing House, 2014. 172 p. (in Russ.).

D E N T I S T R Y


EVALUATING OPTICAL DENSITY OF ALVEOLAR BONE IN PATIENTS WITH DIABETES MELLITUS USING CONE-BEAM COMPUTED TOMOGRAPHY

A B S T R A C T — The study involved an analysis of optical density of the maxillary and mandibular bone tissue based on cone-beam computed tomography data. It has been shown that the structure and bone tissue density depend on the severity of diabetes mellitus and complications. The results of the study revealed a significant decrease in the bone tissue optical density at the tooth necks in people suffering from diabetes mellitus, whereas fewer changes were manifested at the middle third of the dental roots. Minor changes or even an increase in the optical density were observed at the dental root tips.

K E Y W O R D S — diabetes mellitus, dental computed tomography, alveolar bone tissue, optical density.

Natalya Prozorova1 , Roman Fadeev1,2,3 ,Victor Weber1, Marina Chibisova, Natalia Robakidze2, Irina Prozorova1 ,Sofya Sokolova1, Vladimir Shkarin4

1 Yaroslav-The-Wise Novgorod State University, Veliky Novgorod;2 North-Western State Medical University named after I.I. Mechnikov, St. Petersburg;3 St. Petersburg Institute of Dentistry of Postgraduate Education, St. Petersburg; 4 Volgograd State Medical University, Volgograd, Russia

[email protected]

Received 29 April 2021; Received in revised form 3 June 2021; Accepted 7 June 2021

I N T R O D U C T I O NThe past decade has witnessed an increase in the

number of diabetic patients in Russia by more than 1 million people, yet the true prevalence of the disease is 2–3 times as high [1].

Diabetes mellitus is a metabolic disease entail-ing a high level of complications [2–8]. The literature claims [9–11] that violated connections of protein and mineral components, as well as deteriorated trophic tissues in case of diabetes mellitus lead to delayed re-modeling of bone tissue, which is the factor determin-ing its density. Diabetic arthropathies represent a fairly common complication of diabetes mellitus and affect, as has been described in a number of publications, 58% of patients with Type I diabetes and 24% of patients

with Type II diabetes [12]. The endocrine system has a complex effect on the musculoskeletal tissues structure and function, which suggests that either insufficient or excessive production of a particular hormone will lead, sooner or later, to pathological changes in bones, joints and muscles [13–15].

At the same time, apart from changes involving the quantitative features of bone tissue, it is important to investigate the qualitative parameters of bone, in-cluding in patients with diabetes mellitus. In particu-lar, experiments on animals with insulin deficiency followed by a histomorphometric analysis of the obtained data indicate a decrease in the bone tissue de-velopment rate [16–18]. Besides, there was a decrease observed in the length of bone trabeculae, as well as in the periosteal and endocortical surfaces of the osteoid-covered cortical plate. At the same time, a decrease was observed in the number of osteoblasts, and a distur-bance in their function, as well as an increase in the apoptosis rate [19–21].

Diabetes mellitus has been found to be a factor predisposing to the onset and progress of destruc-tive periodontal diseases that lead to the loss of the dentogingival junction [25]. The current concept [24] holds it that in patients suffering from Type II diabetes mellitus, the primary role in the pathogenesis of inflammations affecting the alveolar part belongs to microangiopathies and acidosis due to high blood glucose content. Insulin insufficiency leads, on the one hand, to a decrease in the synthesis of collagen and al-kaline phosphatase by osteoblasts, while the produced substances are involved in the development and miner-alization of the intercellular matrix, and, on the other, to disturbances in the calcium absorption by the small intestine microvilli and its increased excretion from the body with urine. Hypocalcemia, in turn, stimulates the synthesis of parathyroid hormone, which results in thinning of the compact layer and in the bone tissue resorption.

Therefore, diabetes mellitus is accompanied with disturbed metabolic processes in the bone, disrupted functioning of its cellular elements and organic struc-ture, which leads to violated biomechanical proper-ties and, consequently, a greater risk of fractures. The

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/28

https://orcid.org/0000-0002-8546-0991

https://orcid.org/0000-0003-3467-4479

https://orcid.org/0000-0002-4903-0142

https://orcid.org/0000-0001-7158-0282

http://dx.doi.org/10.35630/2199-885X/2021/11/2/28


potentially involved mechanisms that at least offer a partial explanation to the causes behind bone deterio-ration in case of diabetes, include hyperglycemia and microangiopathy. Bone tissue fragility is an effect of insufficient exposure to insulin, and not a complication of diabetes mellitus [23]. This means that the mecha-nisms underlying the developing bone fragility are the same for both types of diabetes. Besides, it has no direct relation to diabetes and can occur long before its clinical manifestations.

Jaws bone tissues differ little from the rest of the skeleton in terms of their chemical composition and structure. However, the alveolar bone has internal restructuring processes going on faster than in other bones of the skeleton. Normally, the height of the alveolar ridge is maintained by the physiological bal-ance between bone formation and resorption, which is regulated not only with systemic, yet also with local factors [26–29].

In modern dentistry, X-ray diagnostics methods make up an integral part of a comprehensive medical ex-amination. Computed tomography (CT) is a relatively new method used to study the chewing system that allows obtaining high-resolution 3D images. Compared to 2D X-rays, 3D digital computed tomography can im-prove significantly diagnostics quality, including differ-ential diagnostics, as well as reduce the risk of errors. All this is due to a higher resolution of the images obtained, as well as to the option, which allows a layer-by-layer examination on the computer screen [14, 22, 30].

AimThe aim of this study was to identify optical den-

sity of the bone tissue in the alveolar parts of the upper and lower jaw in patients with diabetes mellitus using cone-beam computed tomography.

C L I N I C A L P O O L A N D S T U D Y M E T H O D S

The comprehensive study involved 94 patients aged 31–75 (the median age being 53±5, with no gender factor taken into consideration), who were divided into 2 groups. Group 1 included 44 patients with diabetes mellitus (6 patients with Type I diabetes and 38 patients — with Type II diabetes), while Group 2 included 50 patients with no endocrine pathology. A clinical dental examination performed in order to carry out additional diagnostics, prior to orthopedic or surgical treatment, all patients were to undergo a cone-beam computed tomography examination.

The inclusion criteria employed through the study were: age — 18 and above; Type I or Type II diabetes mellitus in the history; lack of dentition issues or the presence of some small (1 to 3 teeth missing)

and medium (4 to 6 teeth missing) defects through the dentition length. The inclusion criteria were: endocrine pathology accompanying diabetes mellitus; chronic diseases in the decompensation stage; onco-logical issues, as well as the following dental diseases identified: pathological tooth abrasion; large dentition defects, periodontitis. The exclusion criterion implied the patient’s refusal to undergo the respective examina-tions.

The examination was performed on a Gendex-GXCB-500 cone-beam computed tomograph using the Icat Vision software. The optical density on the tomograms was estimated using a density window with a side of 3 mm. The measurements within the groups were carried out in the interdental septa of the teeth present in both jaws at the levels of their roots’ mid- and top points, as well as the alveoli upper edges. Each of the measurements was performed three times. The average density value was calculated automatically by the software. The optical density was expressed in Hounsfield units (HU). The central trend and data dispersion were calculated with descriptive statistics methods. The quantitative parameters, depending on the distribution type, were presented as the mean value (M) and the mean square deviation (SD), or, when performing the assessment with nonparametric statistics, as the median (Xmed) and the interquartile QR range within the (LQ 25%÷UQ75%) range.

The analysis of the correspondence between the feature distribution type and the law of normal distribution was performed using the Shapiro-Wilk test. The critical level of difference significance when testing the statistical hypotheses was set at p<0.05. The nonparametric Mann-Whitney method was employed to analyze the differences between the subgroups identified subject to qualitative clinical and diagnostic features, with the Bonferroni correction factor used to estimate the Student’s test values. The statistical processing of the obtained data was carried out using the Statistica 10 software.

S T U D Y O U T C O M E S As shown in Figures 1–6, the most significant

changes affecting bone density could be seen at the tooth necks of the upper and lower jaws. Changes in bone density at the central part of the dental roots were less significant. Patients with this pathology featured an increase in the bone density at the tops of a number of dental roots.

A comparative analysis of the studied values revealed significant differences in the Gauss density in the group of patients, suffering from diabetes mellitus, due to a decrease in the bone tissue optical density, especially in the lower jaw (Tables 1–3).

D E N T I S T R Y


Fig. 1. Comparative characteristics of the maxillary bone tissue optical density by the Gauss density value at the tooth apices

Fig. 2. Comparative characteristics of the mandibular bone tissue optical density by the Gauss den-sity value at the tooth apices

Fig. 3. Comparative characteristics of the maxillary bone tissue optical density by the Gauss density value at the central part of the dental roots

Fig. 4. Comparative characteristics of the mandibular bone tissue optical density by the Gauss density value at the central part of the dental roots

Fig. 5. Comparative characteristics of the maxillary bone tissue optical density by the Gauss density value at the tooth necks Fig. 6. Comparative characteristics of the mandibular bone tissue optical

density by the Gauss density value at the tooth necks

A comparative analysis of optical density para-metric data in the mandibular bone tissue in patients of the main and control groups revealed differences (P<0.05) at the dental root apices: 3.1, 4.1, 4.3, 4.4, 4.6 4.8; at the central part of the dental roots: 3.4, 3.3, 3.2,

3.1, 4.1, 4.2, 4.3, 4.4, 4.6; and almost at all the tooth necks, except tooth 3.8 (Table 4–6).

A nonparametric analysis helped reveal a range of differences in view of the median and the interquartile

D E N T I S T R Y


Table 1. Bone tissue optical density parametric data at the apices of the maxillary dental roots, HU

Tooth no. М±SD m М±SD m t PPatients without diabetes Patients with diabetes1.8 235,2±107,3 87,4 166,7±47,7 32,9 1,626 0,04921.7 193,1±124,6 103,2 479,3±362,1 286,1 -3,807 0,00001.6 284,0±163,8 140,7 418,7±320,1 250,8 -1,517 0,02231.5 302,0±158,6 134,3 566,7±294,9 232,1 -3,426 0,01841.4 252,1±138,5 116,1 449,4±348,9 299,9 -2,427 0,00071.3 255,3±143,1 124,2 395,6±345,3 288,1 -2,067 0,00011.2 187,3±127,3 102,1 238,3±249,0 152,0 -0,932 0,00271.1 218,5±133,0 108,5 291,3±283,5 221,8 -1,230 0,00062.1 227,1±146,2 124,7 196,6±195,9 137,1 0,540 0,20752.2 239,9±191,5 145,7 151,2±75,8 51,4 1,357 0,00952.3 270,9±120,8 98,9 306,3±161,8 129,1 -0,850 0,16382.4 194,1±104,2 85,1 305,7±170,6 131,0 -2,517 0,03972.5 260,6±111,1 87,1 378,1±290,6 208,7 -1,926 0,00012.6 358,2±229,9 196,2 411,8±269,1 200,3 -0,580 0,49612.7 223,7±127,2 101,3 307,9±258,0 210,7 -1,320 0,00662.8 238,9±181,0 148,7 292,5±144,5 129,8 -0,653 0,6504

Tooth no. М±SD m М±SD m t PPatients without diabetes Patients with diabetes1.8 237,0±95,1 68,8 133,6±40,4 30,8 2,781 0,04111.7 308,6±171,5 134,2 333,7±361,1 289,2 -0,281 0,00581.6 434,1±247,0 156,5 329,3±339,4 295,0 0,851 0,26881.5 302,8±106,9 87,2 506,6±282,3 228,4 -3,315 0,00021.4 447,4±184,1 148,3 364,8±211,3 163,5 0,987 0,56511.3 327,7±124,0 95,2 309,3±291,9 229,0 0,317 0,00011.2 434,0±222,2 189,0 189,5±193,5 145,8 3,203 0,69121.1 363,0±244,2 200,1 210,7±298,3 217,1 1,709 0,36302.1 473,6±256,1 227,1 181,1±329,1 193,5 2,982 0,27602.2 385,5±210,5 172,9 174,4±106,8 89,1 2,912 0,04802.3 475,4±265,8 188,7 314,0±242,2 209,5 2,029 0,40722.4 354,5±133,7 103,1 237,9±179,5 137,3 2,228 0,21242.5 470,4±224,4 193,8 304,0±244,2 195,8 0,649 0,16602.6 254,8±115,9 96,4 227,4±195,3 135,2 2,834 0,74792.7 304,5±170,4 143,9 132,4±54,8 44,8 2,712 0,06692.8 237,0±95,1 68,8 264,0±223,7 172,4 0,437 0,3879

Note: Values at P <0.05 are italicized

Table 2. Bone tissue optical density parametric data at the central part of the maxillary dental roots, HU


range (QR). Table 7 contains data describing the dif-ferences in the upper jaw, while the respective data for the lower jaw are presented in Table 8.

The obtained data point at significant changes affecting the optical density of bone tissue in patients with diabetes mellitus versus the control group due

D E N T I S T R Y


Table 3. Bone tissue optical density parametric data at the maxillary tooth necks, HU


Tooth no. М±SD m М±SD m t PPatients without diabetes Patients with diabetes1.8 318,5±132,9 89,5 66,9±33,5 23,8 5,244 0,00101.7 349,7±177,2 140,8 129,7±72,0 56,8 2,978 0,04901.6 400,2±281,6 200,3 176,2±144,1 101,8 1,733 0,19891.5 315,3±106,7 85,2 250,3±149,2 125,1 1,370 0,20101.4 433,5±184,1 143,3 145,2±67,6 53,4 3,753 0,03391.3 350,5±144,2 121,5 174,9±179,4 154,3 3,323 0,32501.2 391,5±163,7 131,7 156,0±166,3 98,2 4,077 0,86401.1 435,8±220,7 174,9 148,5±152,3 92,2 3,896 0,23502.1 430,6±235,4 134,7 130,9±107,1 93,0 5,472 0,47832.2 486,7±197,9 172,0 98,3±79,5 53,6 5,748 0,01052.3 426,4±463,7 245,7 129,4±118,5 85,5 3,655 0,02602.4 488,7±333,3 192,7 115,5±76,0 49,6 5,228 0,00772.5 389,5±236,8 146,7 173,8±174,9 111,9 3,018 0,58002.6 423,3±165,3 134,1 144,7±109,2 80,1 4,780 0,21842.7 281,2±195,6 153,8 101,0±59,6 37,3 2,217 0,01442.8 345,1±126,9 96,9 93,3±41,6 29,2 5,081 0,0113

Table 4. Bone tissue optical density parametric data at the apices of the mandibular dental roots, HU


Tooth no. М±SD m М±SD m t PPatients without diabetes Patients with diabetes3.1 267,7±166,9 138,4 92,3±60,1 40,3 3,088 0,0033123.2 253,3±195,4 149,4 125,2±90,1 71,4 1,914 0,0612673.3 265,2±151,2 121,0 285,5±308,0 251,6 -0,317 0,7523033.4 231,1±128,6 96,0 288,7±255,2 199,8 -1,030 0,3079343.5 268,9±181,4 154,0 426,4±144,7 96,7 -1,855 0,0712303.6 265,4±164,7 138,3 264,3±291,9 185,9 0,012 0,9904073.7 277,1±190,3 163,5 130,8±68,8 45,4 1,681 0,1024743.8 235,2±151,4 120,2 395,6±305,9 249,7 -1,730 0,0965014.8 385,0±171,8 146,9 127,7±115,8 93,0 3,482 0,0014614.7 304,6±198,0 168,3 501,2±371,6 293,8 -1,788 0,0832674.6 474,9±217,5 180,5 139,7±142,7 98,9 3,526 0,0017284.5 215,7±94,4 75,0 289,3±166,8 130,1 -1,566 0,1254714.4 283,4±129,6 109,9 123,0±64,5 46,2 3,778 0,0004284.3 340,3±187,5 152,9 165,6±176,5 113,9 2,888 0,0056034.2 181,4±113,4 89,3 115,4±70,3 57,6 1,672 0,1006684.1 262,7±160,4 131,3 82,3±58,3 42,5 3,304 0,001785

to the results of both parametric and nonparametric analysis.

R E S U L T S A N D D I S C U S S I O NThe analysis of the bone tissue optical density

results in patients with diabetes mellitus, when com-

D E N T I S T R Y


Table 5. Bone tissue optical density parametric data at the central part of the mandibular dental roots, HU

Note: Values at P <0.05 are italicized.

Tooth no. М±SD m М±SD m t PPatients without diabetes Patients with diabetes3.1 487,7±274,1 221,1 135,9±82,0 59,4 3,787 0,0004173.2 453,1±321,6 285,2 101,7±49,3 33,3 3,246 0,0020903.3 472,9±237,2 202,0 207,0±256,6 170,9 3,282 0,0018083.4 412,7±209,5 178,2 212,7±214,9 178,0 2,700 0,0094303.5 346,0±203,9 164,9 195,2±202,4 160,6 1,551 0,1290723.6 330,3±135,2 95,6 323,6±241,6 211,5 0,093 0,9268493.7 351,2±157,3 123,7 302,5±329,3 250,0 0,398 0,6929923.8 249,8±102,3 79,5 348,5±307,5 258,2 -1,292 0,2083384.8 422,6±187,3 156,0 238,4±203,7 139,0 2,002 0,0490204.7 383,1±182,5 137,6 365,2±308,8 224,2 0,183 0,8561314.6 414,3±222,5 164,6 100,5±64,8 52,8 3,368 0,0025514.5 347,2±201,0 149,7 314,8±272,6 219,5 0,346 0,7310234.4 434,8±169,7 132,3 111,7±81,9 55,4 5,823 0,0000004.3 484,4±266,2 192,2 161,0±172,0 117,0 3,970 0,0002184.2 396,9±231,3 176,2 114,6±83,3 62,0 3,590 0,0007424.1 472,0±307,2 243,9 98,3±56,9 51,3 3,608 0,000722

Table 6. Bone tissue optical density parametric data at the mandibular tooth necks, HU

Note: Values at P <0.05 are italicized.

Tooth no. М±SD m М±SD m t PPatients without diabetes Patients with diabetes3.1 344,1±178,8 119,2 86,2±42,9 34,3 4,268 0,0000903.2 349,3±202,3 139,3 94,7±54,2 43,3 3,723 0,0004923.3 360,5±186,1 149,7 98,1±62,2 44,7 4,587 0,0000273.4 314,5±173,0 135,5 141,9±208,1 120,0 2,728 0,0087603.5 357,7±168,5 124,4 139,8±95,0 72,6 2,809 0,0077343.6 348,4±166,8 143,0 153,9±58,7 44,7 2,992 0,0060023.7 337,4±160,2 132,9 144,0±84,0 59,7 2,853 0,0074183.8 239,6±109,3 85,5 154,0±142,8 94,0 1,583 0,1259024.8 355,4±180,8 147,9 91,7±50,5 41,0 3,506 0,0013724.7 369,4±254,6 166,9 111,4±87,3 61,7 2,217 0,0340754.6 435,0±226,4 189,3 96,9±45,4 37,6 3,876 0,0006814.5 496,7±248,8 202,8 155,0±88,6 76,0 3,014 0,0045694.4 469,9±425,6 228,1 115,5±88,4 70,5 2,601 0,0122694.3 489,6±439,4 222,9 116,8±69,2 59,7 2,910 0,0052754.2 298,3±147,5 105,3 77,7±37,0 32,1 4,428 0,0000504.1 441,6±213,9 165,3 76,6±43,4 32,0 5,058 0,000006

pared to the control group, showed that the most sig-nificant changes can be observed at tooth necks. Less significant changes were observed at the middle third

of the dental roots. Minor changes or even an increase in the bone optical density were to be seen at the den-tal root apices. This data is comparable with the data

D E N T I S T R Y


available in the respective Russian literature (Bondar-enko N. N., Balakhontseva E. V., 2012, Nikolayuk V. I., Kabanova A. A., Karpenko E. A., 2015, Chuev V. P. et al., 2017, Khaibullina R. R. et al., 2018).

The upper bone density in the jaw of patients with diabetes mellitus ranged from 151.2 ±75.8 to 566.7±294.9 Hu at the tooth apices; from 132.4±54.8 to 506.6±282.3 Hu — at the central part of the roots, while at the tooth necks it ranged from 66.9±33.5 to 250.3±149.2 Hu. As for the lower jaw bone density, it ranged from 82.3±58.3 Hu to 501.2±371.6 Hu.in at the tooth apices; from 98.3±56.9 Hu to 365.2±308.8 Hu — at the cen-tral part of the roots, and from 76.6±43.4 HU to 155.0±88.6 Hu at the tooth necks.

Thus, there is a direct relationship to be seen between changes affecting bone density at the tooth necks, especially in the molars and incisors, which is explained by deteriorating periodontal trophism, inflammatory issues and a slowdown in bone remod-eling. Changes affecting the area at the tooth apices are less significant. The area at the lateral group of teeth featured an inverse relationship, and here the bone density in individuals with diabetes exceeded that in the control group, which can be explained by the effect of reparative processes and the nature of blood supply in this area.

C O N C L U S I O N1. The bone tissue status of the upper and lower

jaw alveolar parts, as could be seen from cone-beam

Table 7. Characteristics of bone tissue optical density nonparametric data for the upper jaw, HU

Tooth no. Xmed by QR Xmed by QR

Patients without diabetes Patients with diabetesIn the area of the apex of the teeth1.8 268,0 130,5÷302,0 168,0 141,0÷211,01.7 143,0 90,0÷263,0 432,4 187,0÷802,01.6 266,0 124,0÷436,0 419,0 117,3÷641,01.5 291,0 167,0÷421,0 595,0 314,0÷901,01.4 224,0 145,0÷366,0 402,5 123,5÷698,01.3 222,5 118,5÷392,0 208,5 152,0÷640,01.2 155,0 100,0÷267,0 166,0 98,0÷253,01.1 243,5 86,5÷283,0 169,5 83,0÷475,02.2 165,5 115,0÷285,5 148,0 115,0÷172,02.4 167,5 127,0÷257,5 324,0 155,0÷407,02.5 257,0 173,0÷356,5 245,0 128,5÷507,52.7 209,0 109,0÷329,0 184,0 81,0÷617,0 In the area of the central part of the roots of the teeth1.8 229,0 177,0÷266,5 142,0 88,0÷172,01.7 260,5 152,0÷412,0 190,0 47,0÷654,01.5 316,0 222,0÷385,0 498,0 238,0÷824,01.3 301,0 264,0÷407,5 206,0 77,0÷444,02.2 365,5 213,0÷541,0 198,0 86,0÷223,0In the area of the necks of the teeth1.8 299,0 234,0÷380,0 64,0 45,0÷77,01.7 308,5 245,0÷444,0 147,0 66,0÷179,01.4 421,0 329,0÷526,0 157,5 76,0÷205,02.2 504,0 321,0÷631,0 72,0 56,0÷79,02.3 293,0 221,0÷461,0 83,0 50,0÷151,02.4 461,0 326,0÷589,0 95,0 71,0÷120,02.7 229,0 98,0÷361,0 92,5 84,0÷108,02.8 309,0 278,0÷445,0 93,0 63,0÷120,0

D E N T I S T R Y


Table 8. Characteristics of bone tissue optical density nonparametric data for the lower jaw, HU

Tooth no. Xmed by QR Xmed by QRPatients without diabetes Patients with diabetes

In the area of the apex of the teeth3.1 288,5 73,0÷397,0 81,0 56,0÷86,04.8 417,5 214,0÷522,5 89,0 34,0÷211,04.6 489,0 314,5÷652,0 82,5 62,0÷157,04.4 296,0 163,0÷379,0 97,5 84,0÷156,04.3 348,0 212,0÷487,0 112,5 62,0÷160,54.1 290,5 75,0÷362,0 66,0 47,0÷72,0In the area of the central part of the roots of the teeth3.1 516,0 264,0÷667,0 127,0 91,0÷161,03.2 470,0 134,0÷758,0 96,0 85,0÷109,03.3 475,0 248,0÷700,0 89,0 78,0÷292,03.4 445,0 236,0÷548,0 93,5 56,0÷436,04.6 397,5 239,0÷559,5 84,5 51,0÷161,04.4 431,0 329,0÷500,0 83,0 65,0÷140,04.3 477,5 294,0÷632,0 106,0 55,0÷207,04.2 422,0 167,5÷496,0 107,0 51,0÷161,04.1 455,5 254,0÷658,5 64,0 57,0÷146,0In the area of the necks of the teeth3.1 337,5 231,0÷421,0 77,0 51,0÷108,03.2 332,0 224,0÷437,5 82,0 68,0÷126,03.3 320,0 231,0÷456,0 73,0 62,0÷110,03.4 258,0 189,0÷424,0 77,0 36,0÷115,03.5 339,0 246,0÷419,0 115,0 82,0÷176,03.6 296,0 241,0÷512,0 139,0 116,0÷205,03.7 314,0 241,0÷444,0 118,5 109,0÷170,04.8 295,5 225,0÷501,5 79,0 50,0÷145,04.7 301,0 225,0÷450,5 90,0 56,0÷225,04.6 418,5 233,5÷588,0 116,0 60,0÷120,04.5 459,0 322,0÷758,0 201,0 81,0÷225,04.4 366,0 256,0÷491,0 99,5 39,0÷166,04.3 428,0 314,0÷525,5 86,5 71,5÷176,04.2 266,0 193,0÷318,5 95,0 46,0÷100,04.1 450,0 314,0÷612,5 67,0 53,0÷82,0

computed tomography, showed a significant differ-ence between the control group and the patients with diabetes. In case of diabetes, changes in the bone tissue optical density affected both the upper and the lower jaw, and were symmetrical in nature. Certain specific features identified on the right and on left halves may be explained by some peculiarities in chewing.

2. Patients with diabetes had the most significant changes in the bone tissue at the tooth necks of the up-per and lower jaw, while the changes implied a decrease in the optical density.

3. Changes in the bone tissue optical density at the apices as well as at the central part of the dental roots were less pronounced. A decrease in the bone optical density in the patients with diabetes mellitus could be observed at the anterior group of teeth, both in the upper and in the lower jaw. At the molars and premolars of the upper and lower jaws, in the central part of the roots as well as at the dental root apices, there was an increase in the bone optical density.

4. The study outcomes suggest that changes in optical density can be used as an evaluation criterion

D E N T I S T R Y


for assessing the jaw bone tissue status in people with diabetes mellitus. It could be also viewed as an im-portant sign of early detection and forecasting of the progression of periodontal tissue disease.

R E F E R E N C E S1. Dedov I. I., Endocrinology: textbook / I. I. Dedov,

G. A. Melnichenko, V. V. Fadeev – M.: Litterra, 2015. – 416 pp. (In Russ.).

2. Basov A.A., Ivchenko L.G. The role of oxidative stress in the pathogenesis of vascular complications in children with insulinable sugar diabetes // Archiv EuroMedica. 2019. Vol. 9; 1: 136–145. https://doi.org/10.35630/2199-885X/2019/9/1/136

3. Davydov B.N. Clinical and functional approaches to comprehensive treatment of periodontal dis-eases in children with type I diabetes. Parodon-tologiya. 2021;26(1):9–19. (In Russ.) https://doi.org/10.33925/1683-3759-2021-26-1-9-19

4. Davydov B.N. Peculiarities of microcirculation in periodont tissues in children of key age groups suffi-cient type 1 diabetes. Part I. Periodontology, 2019; Vol. 24; 1–24(90): 4–10. DOI: 10.25636/PMP.1.2019.1.1

5. Davydov B.N. Peculiarities of microcirculation in periodont tissues in children of key age groups sufficient type 1 diabetes. Part II. Periodontology. 2019;24(2):108–119. (In Russ.) DOI:10.33925/1683-3759-2019-24-2-108-119

6. Davydov B.N. Modern possibilities of clinical-lab-oratory and x-ray research in pre-clinical diagnostics and prediction of the risk of development of periodon-tal in children with sugar diabetes of the first type. Part I. Periodontology, 2018; Vol. 23; 3–23(88): 4–11. DOI:10.25636/PMP.1.2018.3.1

7. Domenyuk D.A. Contemporary methodological approaches to diagnosing bone tissue disturbances in children with type 1 diabetes. Archiv Euro-Medica, 2018; 8(2): 71–81. DOI:10.35630/2199-885x/2018/8/2/71

8. Domenyuk D.A. Matrix metalloproteinases and their tissue inhibitors in the pathogenesis of peri-odontal diseases in type 1 diabetes mellitus // Archiv EuroMedica. 2019. Vol. 9. № 3. P. 81–90. https://doi.org/10.35630/2199-885X/2019/9/9/3.25

9. Shepelkevich A.P. Morphological changes in bone tissue in type 1 diabetes mellitus / Shepelkevich A.P., Kabak S.L., Rogov Yu.I., Kaban N.S., Lebed O.A. // Military medicine. – 2011 – №4 (21) . – p. 68–73 (In Russ.).

10. Shepelkevich, A. P. Osteoporosis-a complication of diabetes, which is given insufficient attention /A. p. Shepelkevich, O. V. Zhukovskaya, O. A. Shakulya // Medical journal. – 2008. – No. 2. – Pp. 91–95. (In Russ.).

11. Bone mineral density in patients with type 1 and type 2 diabetes / J.T. Tuominen [et al] // Diabetes Care. – 1999. – Vol. 2561, No 22. – P. 1196–2000. DOI: 10.2337/diacare.22.7.1196

12. Bone mineral density measured by dual x-ray absorp-tiometry in Spanish patients with insulindependent diabetes mellitus / М. Muñoz-Torres, et al // Calcif. Tissue Int. – 1996. – 58(5). – Р. 316–319. DOI: 10.1007/BF02509378

13. Holt S.C. Factors in virulence expression and their role in periodontal desease pathogenesis // Crit. Rev. Oral Biol. Med. — 1991. — Vol. 2, No 2. — P. 177–281. DOI: 10.1177/10454411910020020301

14. Yanushevich O. O. Dental morbidity of the popu-lation of Russia. — M.: Moscow state University of medicine, 2008. – 228 pp. (In Russ.).

15. Verbovaya N.I., Kosareva O.V. Mineral density of bone tissue and its metabolism in type 2 diabetes mel-litus in patients of older age groups", 2003. (In Russ.).

16. Kemink, S.A. Osteopenia in insulin-dependent dia-betes mellitus; prevalence and aspects of pathophysi-ology / S.A. Kemink, et al. // J Endocrinol Invest. – 2000. – Vol.23, No 5 – P.295–303. DOI: 10.1007/BF03343726

17. Bone mineral density of both genders in Type 1 diabetes according to bone composition / D.J. Hadjidakis, et al // J. Diabetes Complications. – 2006. – Vol. 20, No 5. – Р. 302–307. DOI: 10.1016/j.jdiacomp.2005.07.006

18. Glucose-induced inhibition of in vitro bone minerali-zation / E. Balint, et al // Bone. – 2001.– Vol. 28, No 1. – P. 21–28. DOI: 10.1016/s8756–3282(00)00426-9

19. Histomorphometry of bone tissue: myths and real possibilities / S. L. Kabak, et al. // Health care. – 2007. – No. 12. – pp. 21–24 (In Russ.).

20. Histomorphometric analysis of diabetic osteopenia in streptozotocin-induced diabetic mice: a possible role of oxidative stress / Y. Hamada [et al] // Bone. – 2007. – Vol. 40, No 5. – P.1408–1414. DOI: 10.1016/j.bone.2006.12.057

21. Histomorphometric evaluation of the recovering effect of human parathyroid hormone (1-34) on bone structure and turnover in streptozotocin-induced diabetic rats / Т. Tsuchida, et al // Calcif. Tissue Int. – 2000. – Vol. 66 No 3. – P. 229–233. DOI: 10.1007/pl00005838

22. Bone and mineral metabolism in BB rats with long-term diabetes / J. Verhaeghe [et al] // Decreased bone turnover and osteoporosis. Diabetes. – 1990. – Vol. 37, No 4. – Р. 477–482. DOI: 10.2337/diab.39.4.477

23. Extracellular glucose influences osteoblast differentia-tion and c-Jun expression / M. Zayzafoon, et al // J. Cell Biochem. – 2000. – Vol. 79, No 2. – P. 301–310. DOI: 10.1002/1097-4644(20001101)79:2<301::aid-jcb130>3.0.co;2-0

24. Osteopenia: a bone disorder associated with diabetes mellitus / V.M. Duarte, et al // J. Bone Mineral Me-tabolism. – 2005. – Vol. 23, No 1. – Р. 58–68. DOI: 10.1007/s00774-004-0542-y

25. Is insulin an anabolic agent in bone? Dissecting the diabetic bone for clues. Am. / K.M. Thrailkill, et al //

D E N T I S T R Y


J. Physiol. Endocrinol. Metab. – 2005. – Vol.289, No 12. –P. E735–E745. doi: 10.1152/ajpendo.00159.2005

26. McCabe, L.R. Understanding the pathology and mechanisms of type I diabetic bone loss / L.R. Mc-Cabe // J. Cell Biochem. – 2007. – Vol. 102, No 6. – P. 1343–1357. DOI: 10.1002/jcb.21573

27. Safarova S. S. bone Remodeling in type 1 diabetes. Bulletin of Siberian medicine. 2018; 17 (3): 115–121. (In Russ.).

28. Grebennikova, T. A., White J. E., L. Y. Rozhinskaya etc. Epigenetic aspects of osteoporosis. Bulletin of the Russian Academy of medical Sciences. 2015; 70 (5): 541–548. (In Russ.). doi.org/10.15690/vramn.v70.i5.1440.

29. Yalochkina T.O. Prevalence and factors of occurrence of low-traumatic fractures in type 2 diabetes mellitus. / Yalochkina T.O., Belaya Zh.E., Rozhinskaya L.Ya., Dzeranova L.K., Antsiferov M.B., Shestakova M.V., Melnichenko G.A. // Achievements of personalized medicine today – The result of practical health care tomorrow. – M. – 2016 (In Russ.).

30. Dmitrienko S.V. Analytical approach within cephalo-metric studies assessment in people with various soma-totypes. Archiv EuroMedica. 2019. Vol. 9; 3: 103–111. https://doi.org/10.35630/2199-885X/2019/9/3.29

D E N T I S T R Y


CLINICAL COURSE OF PERIODONTAL DISEASES IN PATIENTS WITH POSTMENOPAUSAL OSTEOPOROSIS TREATED WITH BISPHOSPHONATES

A B S T R A C T — The aim of the study was to assess the state of the clinical course of periodontal diseases in patients with postmenopausal osteoporosis (OP), depending on the long-term oral administration of various groups of bisphosphonates (BP). The study included 120 women aged 55–65 years with postmenopausal OP for at least three years who took complex antiosteoporetic therapy, including BP in tablet form. The dental examination included an examination of the oral cavity, the study of the pH of the oral fluid, the hygienic state of the mouth (the "Florida Probe" system). According to the results of the study, it was revealed that prolonged treatment of BP in tablets can provoke the development of inflammatory reactions in periodontal tissues. This phenomenon is associated with a shift in the pH of saliva to the acidic side, at which its buffer properties change.

K E Y W O R D S — periodontal disease, pH of mixed saliva, "Florida Probe" system, osteoporosis, bisphosphonates.

Marietta Sukhorukikh¹ , Marina Kozlova¹ ,Ekaterina Gorbatova¹ , Larisa Dzikovitskaya² ,Aleksey Bashtovoy¹

1 Central State Medical Academy of the Department of Presidential Affairs, Moscow;2 Central Research Institute of Dentistry and Maxillofacial Surgery, Moscow, Russia

[email protected]


I N T R O D U C T I O NOsteoporosis (OP) is a group of metabolic diseases

of the skeleton characterized by an imbalance in the balance of the bone remodeling cycle, which leads to a decrease in bone mass, a violation of microarchitecton-ics, followed by an increase in fragility and the develop-ment of fractures [10]. A large increase in this pathology was observed in postmenopausal women [10].

Many studies have been described in the literature indicating that a decrease in bone mineral density contributes to a decrease in the height of the inter-dental bone septum, and as a result, the dentoalveolar attachment. Therefore, postmenopausal OP is a risk factor for periodontal tissue diseases [3, 4].

The first-line drugs in the treatment of postmeno-pausal OP are bisphosphonates (BP), whose pharma-cokinetics vary, which is associated with the character-istics of the chemical structure and reflects the degree of therapeutic doses [8, 11].

The most pronounced antiresorptive effect in comparison with other BP has intravenous adminis-tration of the substance, but at the same time the risk of developing osteonecrosis of the jaws increases [7]. Therefore, oral BP is currently the most commonly prescribed treatment in patients with postmenopausal OP [6]. They have low bioavailability, and also have a number of side effects-primarily on the condition of the upper gastrointestinal tract. [9].

Black D.M., Bauer D.C., Schwartz A.V., Cum-mings S.R., Rosen C.J. (2012) note that in 6-30% of cases, these drugs cause a deviation of the pH of the stomach in the acidic side [9].

At the same time, it is proved that low pH values of mixed saliva cause the development of dental dis-eases [9].

In 2007, A. P. Leus, associated changes in the buffer properties of the oral fluid with the values of acid-base equilibrium. When the pH drops to 6.4, the saliva becomes undersaturated with calcium and inorganic phosphate and turns into demineralizing, which subsequently causes the occurrence of caries of hard tooth tissues [5].

According to a number of authors, with a decrease in pH values, the binding property of saliva proteins with calcium increases, playing a role in the processes of demineralization. When this association is violated, calcium is deposited on the surface of the tooth, forming a supragingival tartar, which is formed by saturating the plaque with calcium phosphate crys-tals, which creates an obvious pathogenic factor for the development of periodontal diseases [1,2].

It was also found that the acidification of the pH of saliva secretion is the result of anaerobic processes, resulting in an increase in the activity of hydrolytic enzymes that break down carbohydrates and protein-ases that ensure the adhesion of plaque and microor-ganisms to enamel. At the same time, the activity of acid-stable proteinases increases by 1.5 times, which is

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/29

https://orcid.org/0000-0001-9531-5870

https://orcid.org/0000-0002-3066-206X

https://orcid.org/0000-0002-7729-7979

https://orcid.org/0000-0002-9899-6672

https://orcid.org/0000-0003-3642-967X

http://dx.doi.org/10.35630/2199-885X/2021/11/2/29


accompanied by the appearance of periodontal tissue inflammation.

However, according to the available literature, there are not enough observations to study the devel-opment of dental diseases in patients with primary OP with long-term use of BP in tablet form. Therefore, the problem of the effect of prolonged antiosteoporetic therapy on the dentoalveolar system becomes urgent.

Aim:to evaluate the clinical course of periodontal diseases in patients with postmenopausal osteoporosis, depend-ing on long-term oral administration of various groups of bisphosphonates.

M A T E R I A L S A N D M E T H O D SThe study was conducted at the Department of

Dentistry of the Central State Medical Academy of the Russian Federation from 2017–2020. The study examines 120 patients with primary OP who were treated and monitored at a dispensary for 10 years in the rheumatology department of the Federal State Budgetary Institution "Polyclinic No. 1"

Inclusion criteria: Inclusion criteria: women aged 55 to 65 years with diagnosed postmenopausal OP, who have been treated with BP (per os) for at least three years.

Non-inclusion criteria: patients with surgi-cal menopause, malignant neoplasms, diseases of the parenchymal organs in the decompensation stage, autoimmune diseases, glucocorticoid therapy, therapy with BP for less than three years, the presence of pathological erasability (II, III degrees) of hard tooth tissues in the oral cavity, complete adentia of the denti-tion, secondary adentia of the teeth without ortho-pedic correction.

The diagnosis of osteoporosis was established on the basis of X-ray densitometry performed on the Lunar device in accordance with the Federal Clinical Guidelines for the Diagnosis and Treatment of Oste-oporosis (2014).

120 patients who were on complex antioste-oporetic therapy (per os), including calcium prepara-tions (1000 mg per day) and vitamin D (800 IU daily), depending on the BP taken, were divided into two groups:

I — 51 women — alendronate (70 mg once a week);II — 69 people — ibandronate (150 mg once a

month);The comparison group consisted of 20 female

volunteers aged 55–65 years without osteoporosis with chronic generalized parodontitis.

All subjects signed informed written consents to participate in the study.

The dental status was assessed after detailed monitoring of screening questionnaires, in which the patients, answering questions in detail, covered their health status, taking various medications during their life, including BP, as well as the presence of complaints from diseases in the oral cavity from the beginning of antiresorptive therapy and after three years of treat-ment.

The clinical stage included an examination of the oral cavity, the study of the pH of the oral fluid, the hygienic state of the mouth (the "Florida Probe" system), as well as orthopantomography on the "Plan-meca ProMax"device.

To indicate the acid-base balance, the mixed saliva was collected in the morning on an empty stom-ach in a sterile Petri dish ("Medpolymer" d=60 mm). Before taking the oral fluid, the mouth was rinsed with distilled water. Using a universal indicator paper ("Lach-Ner"), its pH was determined.

The diagnosis of inflammatory periodontal dis-eases was carried out using the "Florida Probe" system ("Florida Probe" Corporation, USA), which contains a probing device, an encoding device that generated data and, after processing them, the digital parameters were visualized on a computer screen.

Titanium dental probe with a movable tube-cou-pling with a diameter of 0.5 mm, provides a smooth study of clinical parameters with a constant pressure of 20 g/cm2. The program automatically determined the following indicators in absolute values: the presence of plaque, the degree of bleeding gums and the depth of periodontal pockets (with high accuracy up to 0.2 mm).

The examination procedure did not cause the patient any sharp pain and discomfort. The received information was displayed on the monitor screen with audio accompaniment and in the printed version in the form of a graphic periodontal map (Fig. 1).

Fig. 1. a — computer visualization of the periodontal map of the "Florida Probe" system, b — diagnostic probe

a b

D E N T I S T R Y


The statistical analysis of the obtained data was calculated by the methods of variational statistics with the calculation of the Student's t-test and the degree of confidence (p) using a computer and the standard software package STATISTICA 10.0 (StatSoft, Inc., USA).

R E S U L T SMonitoring of screening questionnaires revealed

that the number of visits to the general dentist in-creased in patients with postmenopausal OP, regard-less of the group and frequency of taking BP, after six months of treatment, which is associated with com-plaints of partial absence of teeth and periodic swelling of the gums. 93% of women noticed bleeding gums when eating solid food or while brushing their teeth, 73% — indicated the presence of recurrent caries and loss of fillings after six months of taking antiresorptive drugs.

It should be emphasized that all respondents rarely followed the standard rules for taking BF tablets (taking the drug on an empty stomach with a large amount (200 ml) of water, staying at least 1 hour in an upright position), which can be associated with the occurrence of complications from the gastrointestinal tract in the form of gastroesophageal reflux and the appearance of acidity in the stomach.

In all patients with a diagnosed OP, the pH of mixed saliva shifted to the acidic side, and the digital criteria depended on the frequency of taking BP. With weekly administration of alendronate, the acid-base balance parameter was the lowest (5,2±0,03), in comparison with the group receiving monthly therapy (5,9±0,03). Both coefficients significantly (p<0.05) dif-fered from the control group, where the pH is within neutral values (7.4±0.04).

Examining the hygienic state of periodontal tissues, using the device "Florida Probe", in groups I (al-endronate, 70 mg weekly) and II (ibandronate, 150 mg monthly), soft plaque and hard dental deposits were detected (75% — I, 71% — II). However, the data showed no significant differences between the groups and the control.

Analyzing the data of orthopantomography, all women of group I revealed a significant uneven loss of bone tissue of the interalveolar septa within ½–⅔ of the root length, which corresponded to a severe degree of periodontal disease. Mainly, the highest level of it was observed in the area of the lateral group of teeth of both jaws.

Studying the periodontograms obtained us-ing the "Florida Probe" system, in women who took alendronate weekly (70 mg), the value of the degree of bleeding of the gums during probing was equal to

4.6±0.04. Assessing the depth of periodontal pockets, the highest indicator was noted in this group and cor-responded to the level of 7.5±0.04 (Fig. 2). Both signs were statistically significantly higher than the values obtained in women without OP with chronic general-ized parodontitis.

With a monthly intake of ibandronate (150 mg) all subjects of group II were diagnosed with chronic generalized parodontitis of moderate severity. On orthopantomograms, an uneven decrease in the height of the interalveolar septa by ½ the length of the roots in the area of all teeth was determined.

Examining the indicator of the degree of bleeding of the gums in women who were on monthly treat-ment, its average value was established (3.1±0.03). Against this background, the digital values of the depth of the clinical pocket were — 4.78±0.03. As an example, Fig. 3 shows a periodontal chart of patient B. 61 years old, who took ibandronate for more than 3 years.

D I S C U S S I O NLong-term use of complex antiosteoporetic

therapy reduces bone resorption by suppressing the ac-tivity of osteoclasts, which leads to an increase in bone mineral density, but at the same time, long-term use of BF in tablet form has negative sides: it can provoke the development of an inflammatory reaction in periodon-tal tissues, despite systematic observation by a dentist.

This phenomenon is associated with a shift in the acid-base balance of saliva to the acidic side, at which its buffer properties change.

As a result, the progression of periodontal pathol-ogy was observed. We believe that statistically signifi-cant changes in the periodontal status of patients who took ВF are also associated with an increase in the virulence of the periodontal flora in an acidic environ-ment. All of the above correlates with the authors' data indicating the relationship between the development of dental diseases with a decrease in the pH of the mixed secret and a change in its protective functions [1, 2, 5]

It should be particularly noted that women who received weekly therapy with alendronate (70 mg) were in a more unfavorable situation compared to pa-tients with monthly ibandronate (150 mg). According to our data, the degree of gum bleeding during prob-ing was 1.5 times more pronounced and the depth of periodontal pockets was 1.6 times more pronounced (Fig. 4, 5).

C O N C L U S I O NAccording to the results of the study, the highest

risk of an active course of inflammatory periodontal

D E N T I S T R Y


Fig. 3. Periodontal chart of patient B. 61, treated with ibandronate for more than 3 years

Fig. 4. Comparative assessment of the values of gum bleeding in patients of the control, I and II groups (* — the indicators have statistically significant differences with the data in patients with OP and normal bone mineral density), (p<0.05))

Fig. 5. Comparative assessment of the depth of the periodontal pocket in patients of the control, I and II groups who took BP (* — the indicators have statistically significant differences with the data in patients with OP and normal bone mineral density), (p<0.05))

Fig. 2. Periodontal chart of patient B. 61, treated with ibandronate for more than 3 years

D E N T I S T R Y


pathology is exposed to persons who take weekly tablet forms of alendronate 70 mg. It should be recom-mended that patients visit a dentist every three months in order to register the condition of periodontal tissues for preventive and therapeutic measures.

Important in the interdisciplinary interaction is the mandatory referral of women with postmeno-pausal OP, receiving BF, to the dentist to assess the monitoring of the state of the dental system.

R E F E R E N C E S1. Vavilova, T. P., Medvedev, A. E. Biological

chemistry. Biochemistry of the oral cavity: textbook / Vavilova T.P., Medvedev A.E. – M.: GEOTAR-Media, 2014. – 560 p.

2. Gilmiyarova, F. N. Analytical approaches to the study of indicators of metabolism in the oral fluid: textbook / Edited by F. N. Gilmiyarova. – Moscow: Izvestiya, 2012. – 346 p.

3. Dmitrieva, L. A., Atrushkevich, V. G. Compara-tive assessment of the state of mineral metabolism in patients with chronic generalized periodontitis and systemic osteoporosis. – 2009. – No 1. – pp. 30–33.

4. Zamakhaeva, E. V. Photodensitometric determina-tion of bone density in periodontology: abstract of the dissertation of the Candidate of Medical Sciences, Moscow, 2007, 22 p.

5. Leus, P. A. Preventive communal dentistry / P. A. Leus. – M.: Medical book, 2008 – 444 p.

6. Black, D.M., Bauer, D.C., Schwartz, A.V., Cummings, S.R., Rosen, C.J. Continuing bisphos-phonate treatment for osteoporosis — for whom and for how long? // N Engl J Med. 2012; 366 (22): 2051–2053. DOI: 10.1056/NEJMp120262.

7. Black, D.M., Reid, I.R., Cauley, J.A. et al. The Effect of 6 versus 9 Years of Zoledronic Acid Treatment in Osteoporosis: A Randomized Second Extension to the HORIZON — Pivotal Fracture Trial (PFT) // J. Bone Miner. Res. 2015; 30(5): 934–944. doi: 10.1002/jbmr.2442.

8. Cosman, F., de Beur, S.L., LeBoff, M.S. Clini-cian’s Guide to Prevention and Treatment of Oste-oporosis // Osteoporos. Int. 2014; 25(10): 2359–2381. doi: 10.1007/s00198-014- 2794-2.

9. Higuchi, D., Sugawa, C., Shah, S. et al. Etiol-ogy, treatment, and outcome of esophageal ulcers: a 10-year experience in an urban emer- gency hospital. Gastrointest Surg 2003;7(7):836–42. DOI: 10.1007/s11605-003-0027-7

10. Kanis, J.A., Cooper, C., Rizzoli, R., Regin-ster, J.Y.; Scientific Advisory Board of the European Society for Clinical and Economic Aspects of Oste-oporosis (ESCEO) and the Committees of Scientific Advisors and National Societies of the International Osteoporosis Foundation (IOF).European guidance for the diagnosis and management of osteoporosis

in postmenopausal women. Osteoporos Int. 2019 Jan;30(1):3–44. doi: 10.1007/s00198-018- 4704-5

11. Ro, C., Cooper, O. Bisphosphonate drug holiday: choosing appropriate candidates // Curr. Osteoporos. Rep. 2013; 11(1): 45–51. doi: 10.1007/s11914-012-0129-9.

D E N T I S T R Y


INFERIOR ALVEOLAR NERVE BLOCK BY INJECTION INTO THE PTERYGOMANDIBULAR SPACE USING GUIDING DEVICES: A SYSTEMATIC REVIEW

A B S T R A C T — The aim of our research was to analyze and compare different inferior alveolar nerve blocking techniques and the effectiveness of various guiding devices.Methods: A search was conducted on specialized databases for search and selection of works in which a guiding device for lower alveolar nerve block (IANB) was described. The propriated articles were evaluated and selected in 3 stages for final review based on predefined criteria, followed by a critical evaluation stage. As the research result - various types of IANB guide devices and the results of its using were recorded and analyzed. Results: The systematic review of devices for conducting IANB was done. The success of IANB can be achieved by adjusting the syringe with the anesthetic needle trajectory — it increased the probability of a successful hit in the area of the mandible foramen. Examples of devices in which the syringe is fixed at the moment when the tip of the needle is injected into the medial side of the branch of the lower alveolar nerve are considered. And also, the special techniques based on orientation on the soft tissues are described. However, when creating a device of this type, the following factors must be taken into account: the angle of the needle to the insertion point, the position of the insertion point relative to the anatomical landmarks (taking into account individual characteristics), and the insertion depth. Also, other IANB guiding blockade methods are considered: 3D navigation while local anesthetic injection. Thus, the patient's discomfort, the risk of nerve damage and the risk of unsuccessful mandible anesthesia could be minimized. Conclusion: The advantages and disadvantages of these anesthesia methods, the success rate, and patient comfort were analyzed. Prospects for further research in this area were identified.

K E Y W O R D S — anesthesia, inferior alveolar nerve blocking (IANB), guide device for anesthesia, pterygomandibular space.

Artem Kashtanov1 , Hadi Darawsheh3 ,Yuriy Vasil’ev1,2 , Sameh Ibrahim4 ,Darina Shirokova4 , Ekaterina Shilyaeva4 ,Valeria Brichagina4 , Guseyn Taymurazov4

1 I.M. Sechenov First Moscow State Medical University (Sechenov University), Moscow, Russia;2 Kazan State Medical University, Kazan;3 Dental Clinic “Vin-clinic”, Moscow;4 Peoples’ Friendship University, Moscow, Russia

[email protected]


I N T R O D U C T I O NNowadays patients pay great attention to the oral

cavity health, particularly the teeth health (Ibrahim et al., 2017), (Yuan et al., 2020). But most of them still have a fear of dentist visiting. This is due the pain not only during treatment, but also during an anes-thetic injection (Ushnitsky et al., 2018). According to Iwanaga et al., 2018 and Bhat et al., 2020, the most common mandible local anesthesia method is the inferior alveolar nerve blockage (IANB) (Iwanaga et al., 2018), (Bhat et al., 2020). A study by Rajvanshi, 2016 and Howait and Basunbul, 2019 showed a high efficiency of this method — 85% (Rajvanshi H., Ernest S., Hafsa Effendi H., Afridi S., Chhabra M., 2016), (Howait M., Basunbul, 2019). The authors point out that the anesthesia success is an accurate hit in the pterygoid — maxillary space. The most objective way to increase the IANB success is to determine the injec-tion point taking into account the patient anatomical features (Farhangkhoee et al., 2012), (Petrikas A.Zh. et al, 2020). The injection point is located in the once center on the mucosa formed by the protrusion of the anterior inferior alveolar nerve branch edge (outside) and the medial part of the pterygoid muscle (inside) (Iwanaga et al., 2018). There are some intraoral land-marks described for IANB, which may vary depending on an inverted triangle of the individual's anatomy. It should be noted that according to Al-Moraissi et al., 2021, patients experience less pain during the blockade and it is more successful if using 27-gauge needles (Al-Moraissi et al., 2021).

There are a number of adverse events during the IANB. When the needle is inserted into the medial pterygoid-mandibular fold, the anesthetic may spread to the oropharyngeal tissue and damage the internal pterygoid muscle, which could follow to the develop-ment of mandibular contracture (Rabinovich et al., 2018) (Kuzin et al., 2015). When the needle is inserted into the pterygomandibular fold or into its lateral edge, vascular damage, hematoma formation, anes-thetic entry into the bloodstream, and the ischemic zones on the lower lip and chin skin appearance are possible (Alhindi et al., 2016). An allodynia, paresthe-sia, and dysesthesia are most commonly observed

D E N T I S T R Y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/30

https://orcid.org/0000-0002-9648-1720

https://orcid.org/0000-0002-1899-1282

https://orcid.org/0000-0003-3541-6068

https://orcid.org/0000-0002-6233-9929

https://orcid.org/0000-0002-1309-8516

https://orcid.org/0000-0003-1136-1795

https://orcid.org/0000-0002-4433-4526

https://orcid.org/0000-0003-0454-4091

http://dx.doi.org/10.35630/2199-885X/2021/11/2/30


(Ahmad, 2018). This accounts for the need to determine the precision of injection points and the need of a guiding device. (Farhangkhoee et al., 2012). There are some examples of the such devices implementation in the literature but there is no single solution that meets the request for a concise form, ease of use and manufacturer.

M A T E R I A L S A N D M E T H O D SA systematic review of several databases was carried out, with the

studies selection (Scheme 1).

Scheme 1. Methodology for selecting studies for meta-analysis. The original version is taken from the site: http://www.prisma-statement.org/).

R E S U L T S A N D D I S C U S S I O N

The results obtained during the systematic review are presented in Table 1.

Besides, none of the listed devices has a full complete set of necessary functions:

— possibility of a single-uwse;— adjustment for the patient’s

anatomy;— secure attachment of the

syringe;— usage of both sides;— additional maintenance of

the patient's mouth sufficiently open for a dentist full-scale view;

— freeing of the one of the dentist's hands;

— adjustment for the speed and volume of the anesthetic supply

It should be noted that creat-ing of such device should be based on previous studies and existing models. Also it is necessary to take into account such factors as: the needle angle to the injection point, the position of the injection point relative to anatomical landmarks (taking into account individual characteristics) and the depth of the injection. So the guiding devices for the IANB must meet most of the previously named points to ensure not only the success of the blockade, but also the comfort of the dentist and the patient.

C O N C L U S I O NThis systematic review has

shown that using various guiding devices increases the IANB suc-cess. Such devices can help avoid allodynia, prolonged anesthesia, paresthesia, dysesthesia and many other IANB complications. Also it was shown that there is no one point of view on the criteria for evaluating the advantages and disadvantages of guiding devices, so their further development is needed.

D E N T I S T R Y


Tabl

e 1. D

escri

ption

of ex

isting

devic

es fo

r guid

ed in

ferior

alve

olar n

erve

bloc

kage

(IAN

B)

Auth

orAn

esth

esia

type

Devic

e des

crip-

tion

Targ

et in

jectio

n poin

tDe

vice l

ockin

g poin

tsEff

ectiv

enes

sAd

vant

ages

Disa

dvan

tage

sNo

tes

Egor

ov P.

M.,

1982

(Ego

rov P

.M.,

1982

)

By G

ow-

Gate

s

Angu

lator

for

infer

ior al

veola

r ne

rve b

lock

“The

inte

rmus

cular

tri

angle

»

The m

andib

le br

anch

oute

r su

rface

, at t

he le

vel o

f the

man

-dib

ular f

oram

en1 p

atien

tTh

e inj

ectio

n poin

t was

ob

jectifi

ed

Ther

e is n

o sign

ifica

nt

simpli

ficat

ion in

the

anes

thes

ia pr

oced

ure

and t

here

is no

devic

e fix

ing po

ssibil

ity

-

Jofré

and

Mün

zenm

ayer,

19

98(Jo

fré &

M

ünze

nmay

er,

1998

)

By G

ow-

Gate

s

Angu

lator

for

infer

ior al

veola

r ne

rve b

lock

The p

oint i

s dist

al to

th

e sec

ond m

olar o

f the

m

axilla

, slig

htly

below

the

mes

iopala

te pr

otru

sion.

The e

ar pa

rt is

inser

ted i

nto t

he

lower

bord

er of

the t

ragu

s. The

de

vice i

tself i

s orie

nted

in a

plane

fro

m th

e low

er bo

rder

of th

e tra

gus t

hrou

gh th

e cor

ners

of th

e m

outh

. Afte

r the

plan

e is a

ligne

d, th

e syr

inge i

s plac

ed in

the d

evice

an

d the

injec

tion i

s mad

e in t

he

desig

nate

d plan

e.

97,5%

(39

patie

nts a

t all)

The i

njec

tion p

lane

was o

bjecti

fied,

and

objec

tive r

efere

nce

point

s wer

e sele

cted

for a

ll pat

ients

Ther

e is n

o adju

stmen

t fo

r: inj

ectio

n ang

le,

dept

h an

d heig

ht.Th

ere

is no

a co

nven

ient w

ay

to ho

ld th

e dev

ice.

One o

f the

first

objec

tive o

ut-o

f-mou

th

landm

arks

for t

he

block

ade w

as se

lecte

d. It

beca

me p

ossib

le to

ob

jectif

y the

injec

tion

point

.

Tiol A

., 200

1(T

iol A.

, 200

1)

IANB

by

Halst

ed

A dev

ice fo

r ta

rget

ing so

ft tis

sues

. The

m

etho

dolog

y has

be

en im

prov

ed

by Th

anga

velu

et al

., 201

2. (T

hang

avelu

et

al., 2

012)

On Ti

ol A.

, 200

1 met

hod

( Tio

l A., 2

001)

, - d

eepe

ning

of th

e pte

rygo

id lig

amen

t.On

Than

gave

lu et

al., 2

012,

met

hod (

Than

gave

lu et

al.

, 201

2) -

from

6 to

8 m

m ab

ove t

he m

idpoin

t be

twee

n the

uppe

r and

low

er oc

clusa

l plan

es an

d th

e cor

onal

notch

and f

rom

8 t

o 10 m

m po

sterio

rly fr

om

the a

nter

ior bo

rder

of th

e br

anch

On Ti

ol A.

, 200

1 met

hod

( Tiol

A.,

2001

) – m

etal

rod.

It’s th

e edg

e co

incide

s with

the r

eces

s at t

he

pter

ygoid

ligam

ent,

the g

uide

is pa

ralle

l to t

he m

andib

ular o

c-clu

sal p

lane (

Tiol e

t al.,

2010

)

On Ti

ol A.

, 200

1 m

etho

d - (

Tiol A

., 200

1)

met

hod -

94%

( Tio

l A., 2

001)

On Th

anga

velu

et al

., 201

2, m

etho

d (T

hang

avelu

et

al., 2

012)

–

95%

(476

pa

tient

)

On Ti

ol A.

, 200

1 m

etho

d ( Ti

ol A.

, 200

1),

- a co

nven

ient a

nd sa

fe m

etho

d of a

nesth

esia

when

targ

eting

soft

tissu

es.

On Th

anga

velu

et

al., 2

012,

met

hod

(Tha

ngav

elu et

al.,

2012

) – no

addit

ional

devic

es fo

r tar

getin

g th

e tar

get p

oint

The T

iol A.

, 200

1 m

etho

d (T

iol A.

, 200

1):

unsa

fety m

etho

d and

sm

all ev

idenc

e bas

e.Th

e Tha

ngav

elu et

al.

met

hood

(Tha

ngav

elu

et al

., 201

2): –

in-

creas

ed ris

k of in

jury t

o th

e doc

tor. T

he ab

senc

e of

objec

tive a

nd fix

ed

anat

omica

l land

mar

ks

in bo

th ca

ses

The s

uitab

le m

etho

d fo

r tea

ching

stud

ents

to pe

rform

intra

oral

block

anes

thes

ia of

the

infer

ior al

veola

r ner

ve

D E N T I S T R Y

126 | a r c h i v e u r o m e d i c a | 2 0 2 1 | v o l . 1 1 | n u m . 2 |Au

thor

Anes

thes

ia ty

peDe

vice d

escri

p-tio

nTa

rget

injec

tion p

oint

Devic

e loc

king p

oints

Effec

tiven

ess

Adva

ntag

esDi

sadv

anta

ges

Note

s

Ushn

itsky

I.D.,

Chak

hov A

.A.,

2020

(Ush

nitsk

y, I.

D. Ch

akho

v, 20

20)

IANB

by

Halst

ed

Asse

mbly

pack

-ag

e for

injec

tion

point

posit

ion

and t

he de

pth

dete

rmini

ng

It is

possi

ble to

dete

rmine

inj

ectio

ns pa

ram

eter

s dur

-ing

anes

thes

ia ind

ividu

ally

for e

ach p

atien

t

On th

e man

dible

bran

ch po

sterio

r ed

ge in

the a

rea o

f the

grea

test

conc

avity

and fi

x the

mea

surin

g pa

rt of

the s

et w

ith th

e midd

le fin

ger. T

hen t

he ro

d of t

his pa

rt is

fixed

in th

e ora

l cav

ity un

til it

re

ache

s the

stop

in th

e ant

erior

ed

ge of

the l

ower

jaw.

Acco

rding

to

the o

btain

ed m

easu

rem

ents,

th

e dep

th of

imm

ersio

n of t

he

need

le is

dete

rmine

d. Th

en th

e 2n

d par

t of t

he de

vice i

s plac

ed

in th

e ora

l cav

ity un

til it

stop

s at

the m

andib

le fro

nt ed

ge an

d an

injec

tion i

s mad

e alon

g the

guide

ar

ches

.

1 pat

ient

The a

bility

to de

ter-

mine

the d

epth

of

the n

eedle

injec

tion

indivi

duall

y for

each

pa

tient

.

A sm

all am

ount

of da

ta

is ta

ken i

nto a

ccoun

t in

the i

nven

tion,

the

anes

thes

ia its

elf is

pro-

vided

with

out o

bjecti

ve

guide

lines

, the

re is

no

possi

bility

of si

mult

ane-

ous a

djustm

ent o

f all

nece

ssary

para

met

ers,

the l

ack o

f reli

able

fixa-

tion o

f the

devic

e.

It ca

n be u

sed f

or m

en-

tal a

nesth

esia

(Чах

ов

А.А.

, 201

9)

Zand

i and

Se

yedz

adeh

Sa

boun

chi,

2008

(Zan

di &

Seye

dzad

eh

Sabo

unch

i, 20

08)

By G

ow-

Gate

s

Angu

lator

for

infer

ior al

veola

r ne

rve b

lock

Dista

l to t

he m

axilla

ry

seco

nd m

olar a

t the

leve

l of

the m

esiop

alaty

l plan

e.

The e

ar pa

rt is

inser

ted i

nto t

he

lower

bord

er of

the t

ragu

s. The

de

vice i

tself i

s orie

nted

in a

plane

fro

m th

e low

er bo

rder

of th

e tra

gus t

hrou

gh th

e cor

ners

of th

e m

outh

. Afte

r the

plan

e is a

ligne

d, th

e syr

inge i

s plac

ed in

the d

evice

an

d the

injec

tion i

s mad

e in t

he

desig

nate

d plan

e.

93.3%

(45

patie

nts a

t all)

Ease

of ho

lding

the

devic

e, ob

jectifi

ed in

-jec

tion p

lane,

selec

ted

objec

tive r

efere

nce

point

s for

all p

atien

ts

Ther

e is n

o adju

stmen

t fo

r: inj

ectio

n ang

le,

dept

h an

d heig

ht-

Cailli

eux N

., 20

09(C

aillie

ux N

., 20

09)

By G

ow-

Gate

s

The i

njec

tion

sagit

tal p

lane

fixat

ion al

ong t

he

man

dible

bran

ch.

The i

njec

tion

point

selec

tion i

n ac

cord

ance

to th

e sy

ringe

cylin

der

mos

t lat

eral

posit

ion in

that

pla

ne.

The m

ost l

ater

al an

d high

ly loc

ated

in th

e ora

l cav

ity,

afte

r the

devic

e plac

ing.

The e

nds o

f the

devic

e clam

p the

m

andib

ular b

ranc

h bet

ween

its

ante

rior a

nd po

sterio

r edg

es. T

he

injec

tion p

lane i

s sele

cted a

ccord

-ing

to th

e mos

t lat

eral

posit

ion of

th

e syr

inge.

For t

he in

jectio

n, th

e hig

hest

possi

ble po

int in

the o

ral

cavit

y is s

electe

d

No da

taTh

e eas

ing na

vigat

ion

in th

e ora

l cav

ity

Ther

e is n

o reg

ulatio

n fo

r the

patie

nt an

atom

i-ca

l feat

ures

, the

re is

no

regu

lation

of th

e nee

dle

pene

tratio

n dep

th

into s

oft t

issue

s, it

is ne

cessa

ry to

use d

octo

r bo

th ha

nds,

also d

evise

is

not s

table

, the

re is

no

regu

lation

in he

ight

Ther

e is a

prim

ary p

os-

sibilit

y of a

djustm

ent t

o th

e pat

ient i

ndivi

dual

char

acte

ristic

s

Cailli

eux,

2017

(Cail

lieux

, 20

17)

By G

ow-

Gate

s

Angu

lator

for

infer

ior al

veola

r ne

rve b

lock

From

7 to

25 m

m (o

n ave

r-ag

e fro

m 7

to 14

mm

) fro

m

the m

andib

le po

sterio

r ed

ge at

an an

gle of

incli

na-

tion f

rom

53° t

o 62°

(on

aver

age 3

2°)

Fixat

ion be

hind t

he m

andib

le po

sterio

r edg

e, wi

th a

max

imum

lag

of 25

mm

from

that

.

91,6%

A sta

ble an

gle of

incli

-na

tion o

f the

syrin

ge to

th

e inj

ectio

n site

was

cre

ated

, the

syrin

ge

was fi

xed,

the c

alcula

-tio

ns ar

e bas

ed on

the

patie

nts C

T sca

ns da

ta

objec

tive a

nalys

is.

Ther

e is n

o reg

ulatio

n fo

r the

patie

nt an

atom

i-ca

l feat

ures

, the

re is

no

regu

lation

of th

e nee

dle

pene

tratio

n dep

th

into s

oft t

issue

s, it

is ne

cessa

ry to

use d

octo

r bo

th ha

nds,

also d

evise

is

not s

table

, the

re is

no

regu

lation

in he

ight

A num

ber o

f par

am-

eter

s wer

e tak

en in

to

acco

unt:

the i

njec

tion

angle

, the

oral

cav-

ity pa

ram

eter

s in t

he

syrin

ge pa

ssing

plan

e, an

d the

syrin

ge w

as

fixed

in th

e app

ropr

iate

posit

ion

D E N T I S T R Y


Auth

orAn

esth

esia

type

Devic

e des

crip-

tion

Targ

et in

jectio

n poin

tDe

vice l

ockin

g poin

tsEff

ectiv

enes

sAd

vant

ages

Disa

dvan

tage

sNo

tes

Won

and

Kang

, 201

7(W

on &

Kang

, 20

17)

IANB

by

Halst

ed

Supe

rimpo

sing

a CT s

can o

n the

pa

tient

’s ora

l cav

-ity

real

phot

o

Dire

ct or

ienta

tion t

o th

e poin

t of e

ntry

of th

e inf

erior

alve

olar n

erve

into

th

e man

dible

(ach

ieved

by

supe

rimpo

sing a

CT sc

an on

an

intra

oral

phot

o)

-No

data

Выхо

д обе

збол

иван

ия

в AR п

рост

ранс

тво,

разр

абот

ан пр

оток

ол

для п

осле

дующ

их

иссл

едов

аний

Ther

e is n

o sim

plific

a-tio

n in t

he a

nesth

esia

proc

edur

e.

This

is th

e con

venie

nt

met

hod f

or te

achin

g de

ntal

stude

nts t

he

IANB

met

hod.

But i

t is

nece

ssary

to ex

pose

pa

tient

s to e

xces

sive

radia

tion e

xpos

ure a

nd

s to t

rain

docto

rs to

wo

rk in

appli

catio

ns

for p

roce

ssing

DICO

M

imag

es.

Chak

hov A

.A.

Ushn

itsky

I.D.,

2019

(A.A

. Cha

khov

, Us

hnits

ky,

2019

)

By G

ow-

Gate

s

Angu

lator

for

infer

ior al

veola

r ne

rve b

lock

The l

ater

al ed

ge of

the

pter

ygoid

-max

illary

depr

es-

sion,

imm

ediat

ely m

edial

to

the m

edial

bund

le of

the

tem

pora

l mus

cle te

ndon

. Th

e heig

ht of

the i

njec

tion

point

is de

term

ined b

y the

loc

ation

of th

e nee

dle ti

p im

med

iately

below

the

med

ial-li

ngua

l (m

edial

-pa

latal)

max

illary

seco

nd

mola

r tub

ercle

.

The o

uter

ring i

s ins

talle

d in t

he

area

of th

e out

er su

rface

of th

e co

ndyla

r pro

cess.

The d

irecti

on of

th

e inj

ectio

n is c

arrie

d out

in su

ch

a way

that

the t

arge

t mov

es to

th

e spe

cified

point

.

No da

ta

The a

dditi

onal

refer

ence

point

for

anes

thes

ia ha

s bee

n fo

und,

and a

step

has

been

take

n to c

reat

e a

one-

piece

devic

e with

on

ly 1 d

octo

r’s ha

nd

occu

pied.

Ther

e is n

o suffi

cient

jus

tifica

tion f

or th

e use

of

the t

echn

ique,

ther

e is

no w

ay to

objec

tify

the i

njec

tion p

oint,

ther

e is n

o way

to

regu

late t

he in

jectio

n po

int in

heigh

t

-

Jund

t et a

l., 20

20(Ju

ndt e

t al.,

2020

)

By G

ow-

Gate

s

Indiv

idual

mou

thgu

ard f

or

each

patie

nt,

mad

e acco

rding

to

the p

atien

t’s

CT sc

an da

ta

From

3.8 t

o 10 m

m ab

ove

the o

cclus

al pla

ne, 3

mm

m

edial

and 6

mm

high

er

from

the p

oint o

f ent

ry of

th

e inf

erior

alve

olar n

erve

-No

data

Pote

ntial

ly hig

h cha

nce

of su

ccess

of an

esth

esia

due t

o a pe

rsona

lized

ap

proa

ch (t

he ab

ility

to ta

ke in

to ac

coun

t ad

dition

al da

ta)

Exce

ssive

radia

tion

expo

sure

of th

e pat

ient,

high c

ost a

nd lo

ng-te

rm

man

ufac

turin

g of t

he

devic

e

Abilit

y to a

ccoun

t for

m

ore d

ata

Kojim

a et a

l., 20

20Ko

jima e

t al.,

2020

a(K

ojim

a, Se

ndo,

et

al., 2

020)

, (K

ojim

a, M

urou

chi, e

t al.

, 202

0)

IANB

by

Halst

ed

Ultra

soun

d m

anag

emen

t of

anes

thes

iaSt

anda

rd fo

r IAN

B-

1 pat

ient

(Koji

ma,

Send

o, et

al., 2

020)

28 pa

tient

s (K

ojim

a, M

urou

chi, e

t al.

, 202

0)

Easy

to pe

rform

in-

hosp

ital c

ondu

cting

an

esth

esia,

desig

ned

for p

osto

pera

tive a

nd

reha

bilita

tion

The m

etho

d is d

ifficu

lt to

imple

men

t in t

he

cond

ition

s of o

utpa

tient

ad

miss

ion, t

here

is no

ob

jectifi

catio

n of t

he in

-jec

tion p

oint,

ther

e is n

o sig

nifica

nt si

mpli

fica-

tion o

f the

bloc

kade

, it

is ne

cessa

ry to

invo

lve a

spec

ialist

in ul

traso

und

diagn

ostic

s.

-

D E N T I S T R Y


R E F E R E N C E S1. A., Tiol. (2001). Técnica Innovadora Para El Blo-

queo Regional Mandibular Con El Uso De Una Guía Metálica. Practica Odontológica, 22(6), 7–14.

2. A.A. Chakhov, Ushnitsky, I. D. (2019). A device for performing mandibular anesthesia according to Gow-Gates (Patent No. RU 189949 U1).

3. A.A. Chakhov, Ushnitsky, I. D., & Isakov, L. O. (2019). The modified mental anesthesia by malamed technique with Ushnitsky-Chahov’s device. Endodontics Today, 17 (4), 12–15. https://doi.org/10.36377/1683-2981-2019-17-4-12-15

4. A.Zh. Petrikas, E.V. Chestnyh, K.V. Kulikova, I. O. L. (2020). Topographic and anatomical justifi-cation of conduction anesthesia for inferior alveolar nerve block. Tver Medical Journal, 3, 69–78.

5. Ahmad, M. (2018). The Anatomical Nature of Dental Paresthesia: A Quick Review. The Open Dentistry Journal, 12(1), 155–159. https://doi.org/10.2174/1874210601812010155

6. Al-Moraissi, E. A., Al-Selwi, A. M., & Al-Zendani, E. A. (2021). Do length and gauge of dental needle affect success in performing an inferior alveolar nerve block during extraction of adult man-dibular molars? A prospective, randomized observer-blind, clinical trial. In Clinical Oral Investigations. https://doi.org/10.1007/s00784-021-03796-w

7. Alhindi, M., Rashed, B., & Alotaibi, N. (2016). Failure rate of inferior alveolar nerve block among den-tal students and interns. In Saudi Medical Journal (Vol. 37, Issue 1, pp. 84–89). https://doi.org/10.15537/smj.2016.1.13278

8. Bhat, P., Chanu, H. T., Radhakrishna, S., Ashok Kumar, K. R., Marimallappa, T. R., & Ravikumar, R. (2020). Comparison of Clinical Efficacy of Classical Inferior Alveolar Nerve Block and Vazirani–Akinosi Technique in Bilateral Mandibular Premolar Teeth Removal: A Split-Mouth Randomized Study. In Journal of Maxillofacial and Oral Surgery. https://doi.org/10.1007/s12663-020-01374-3

9. Caillieux, N. (2017). Guiding Device for Dental Anesthesia (Patent No. US Patent 0028143 A1).

10. Egorov P.M., E. I. P. (1982). Device for mandible analgesia (Patent No. SU 984480 A1).

11. Farhangkhoee, H., Lalonde, J., & Lalonde, D. H. (2012). Teaching medical students and residents how to inject local anesthesia almost painlessly. In Canadian Journal of Plastic Sur-gery (Vol. 20, Issue 3, pp. 169–172). https://doi.org/10.1177/229255031202000315

12. Howait M., Basunbul, G. I. (2019). Prevalence of Failed Inferior Alveolar Nerve Block (IANB) in Achieving Pulpal Anaesthesia in Mandibular Molars with Symptomatic Irreversible Pulpitis. In Egyptian Dental Journal (Vol. 65, Issue 1, pp. 771–776). https://doi.org/10.21608/edj.2019.72859

13. Ibrahim, H., Lyons, K. M., Armfield, J. M., & Thomson, W. M. (2017). Performance of the Index

of Dental Anxiety and Fear in a population-based sample of adults. In Australian Dental Journal (Vol. 62, Issue 4, pp. 478–484). https://doi.org/10.1111/adj.12541

14. Iwanaga, J., Choi, P. J., Vetter, M., Patel, M., Kikuta, S., Oskouian, R. J., & Tubbs, R. S. (2018). Anatomical Study of the Lingual Nerve and Inferior Alveolar Nerve in the Pterygomandibular Space: Complications of the Inferior Alveolar Nerve Block. Cureus. https://doi.org/10.7759/cureus.3109

15. Jofré, J., & Münzenmayer, C. (1998). Design and preliminary evaluation of an extraoral Gow-Gates guiding device. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontics, 85(6), 661–664. https://doi.org/10.1016/S1079-2104(98)90032-4

16. Jundt, J. S., Chow, C. C., & Couey, M. (2020). Computed tomography-guided 3D printed patient-specific regional anesthesia. Journal of Dental Anesthesia and Pain Medicine, 20(5), 325. https://doi.org/10.17245/jdapm.2020.20.5.325

17. Kojima, Y., Murouchi, T., Akiba, M., & Oka, T. (2020). Ultrasound-guided inferior alveolar nerve block for postoperative analgesia after mandibular sequestrectomy: A single-center retrospective study. Journal of Clinical Anesthesia, 60(August), 39–40. https://doi.org/10.1016/j.jclinane.2019.08.026

18. Kojima, Y., Sendo, R., Ohno, S., & Sugimura, M. (2020). Ultrasound-guided inferior alveolar nerve block for trismus during dental treatment: a case report. JA Clinical Reports, 6(1), 20–23. https://doi.org/10.1186/s40981-020-00400-0

19. Kuzin, A. V., Semkin, V. A., & Dydykin, S. S. (2015). Scientific anatomical rationalefor lower teeth local anesthesia. Stomatologiya, 94(4), 35. https://doi.org/10.17116/stomat201594435-38

20. N., Caillieux. (2009). Device for guiding the syringe of a user so as to enable the injection of the analgesic solution of the former as close to the man-dibular foramen as possible (Patent No. EU Patent EP2429618B1).

21. Rabinovich, S. A., Vasil’ev, Y. L., & Kuzin, A. N. (2018). Anatomic rationale for clinical efficacy of intraosseous mental nerve anesthesia. Stoma-tologiya, 97(2), 41. https://doi.org/10.17116/sto-mat201897241-43

22. Rajvanshi H., Ernest S., Hafsa Effendi H., Afridi S., Chhabra M., N. K. N. (2016). Failure of inferior alveolar nerve block ( ianb ) and techniques to avoid it. European Journal of Biomedical and Pharma-ceutical Sciences, 3(9), 207–210.

23. Thangavelu, K., Kannan, R., & Kumar, Ns. (2012). Inferior alveolar nerve block: Alternative technique. Anesthesia: Essays and Researches, 6(1), 53. https://doi.org/10.4103/0259-1162.103375

24. Tiol, M., Realizar, P., & Dentales, E. (2010). Luz de maría lazo herrera.

D E N T I S T R Y


25. Ushnitsky, I. D. Chakhov, A. A. (2020). Method of mandibular anesthesia (Patent No. RU 2727580 C1).

26. Ushnitsky, I. D., Chakhov, A. A., Vinokurov, M. M., Yurkevich, A. V., Savvina, I. L., & Niki-tin, Y. G. (2018). Modern concept of pain syndrome and dental anxiety pathophysiology and prevention. Stomatologiya, 97(6), 67. https://doi.org/10.17116/stomat20189706167

27. Won, Y.-J., & Kang, S.-H. (2017). Application of augmented reality for inferior alveolar nerve block anesthesia: A technical note. Journal of Dental An-esthesia and Pain Medicine, 17(2), 129. https://doi.org/10.17245/jdapm.2017.17.2.129

28. Yuan, S., Freeman, R., Hill, K., Newton, T., & Humphris, G. (2020). Communication, Trust and Dental Anxiety: A Person-Centred Approach for Den-tal Attendance Behaviours. Dentistry Journal, 8(4), 118. https://doi.org/10.3390/dj8040118

29. Zandi, M., & Seyedzadeh Sabounchi, S. (2008). Design and development of a device for facilitation of Gow-Gates mandibular block and evaluation of its ef-ficacy. Oral and Maxillofacial Surgery, 12(3), 149–153. https://doi.org/10.1007/s10006-008-0126-4

D E N T I S T R Y


CLINICAL AND ANATOMICAL REQUIREMENTS AND INDICATIONS FOR USING A NOVEL MULTIFUNCTIONAL GNATIC DEVICE

A b s t r A c t — Knowledge of the clinical anatomy of the masticatory muscles contributes to the understanding of the etiopathogenetic and physiological aspects of the development of hypertonicity of the masticatory muscles. Hypertonicity of the masticatory muscles is a disease that leads to a number of other dental problems, such as abnormal abrasion, gum recession, trauma to the oral mucosa, wedge-shaped defects. We have developed and patented a pneumatic expander. It is feasible to treat hypertonicity of the masticatory muscles in a complex — myogymnastic exercises on the background of the use of the patented gnatic device. This paper presents clinical and anatomical requirements for the gnathic device as an uncoupling device in the treatment of both teeth and muscle relaxant.

K e y w o r d s — hypertonicity, masticatory muscles, pneumatic trainer, myorelaxant.

Yuliya Makedonova1,2 , Alexander Vorobyev1,2 ,Anna Osyko1 , Alexey Mikhalchenko1 ,Alexander Alexandrov1 , Anastasia Pavlova-Adamovich1

1 Volgograd State Medical University, Volgograd;2 Volgograd Medical Research Center, Volgograd, Russia

[email protected]

Received 24 April 2021; Received in revised form 28 May 2021; Accepted 4 June 2021

in turn irritates pain receptors, causing aggravation of pain symptoms [2, 11]. The emergence of a painful effect is a stressor agent, which leads to a more forced muscle spasm [10], against the background of the production of adrenaline. In parallel with this, there is a decrease in blood flow to the muscles, the formation of edema due to an increase in intercellular fluid and the appearance of painful muscle seals. The contracted muscle fibers lose their ability to function normally, which consists primarily of a partial loss of the chew-ing function [9] and the occurrence of other dental problems. Thus, a vicious circle is formed, which determines the duration of the course of the pathologi-cal process and its chronicity [13]. Based on this, we came to the conclusion that the place of the primary therapeutic effect in Hypertonicity of the masticatory should be the masticatory muscles.

The objective is to develop clinical and anatomical requirements for the use of a multifunctional gnathic device stimulating the masticatory muscles.

M A T E R I A L S A N D M E T H O D SOur proposed invention relates to medicine,

namely to dentistry, and is intended for use in gnathic dentistry, as well as for separating the dentition during medical procedures in the oral cavity [12].

The pneumatic expander device has several ef-fects, which is the clinical success. The functionality of the pneumatic expander-expander when creating an increased internal pressure in the bite block allows the device to be used as a fixer for the volume of mouth opening during various medical procedures in the oral cavity. The possibility of creating pressure in the cavity of the device makes it possible to controllably determine the degree of opening of the mouth, and, accordingly, the degree of stretching of the masticatory muscles, which is necessary in the treatment of patients with hypertonicity of the masticatory muscles. At the same time, the elasticity of the apparatus allows to use it as a stimulator counteracting muscle tissue atrophy. During the treatment and rehabilitation of patients, it is possible to use the apparatus in conjunction with a complex of myogymnastics to increase its effectiveness and reduce the duration of rehabilitation [12].

I N T R O D U C T I O NHypertonicity of the masticatory muscles is

often the cause of a number of pathological symptoms that lead not only to dental problems, but also other diseases (headache, migraine, stomalgia) [14]. Hyper-tonicity of the masticatory can arise under the influ-ence of both general and local factors, while there is a two-way inverse relationship between the underlying factors and vice versa [3–8]. The impact on the group of masticatory muscles of the following factors: 1) General factors — impaired posture, neurogenic fac-tors, autoimmune factors, traumatic factors; 2) Local factors — bite pathology, diseases of the temporoman-dibular joint, iatrogenic factor, bad habits. At the same time, there is an indirect inverse relationship between local and general factors [1]. The consequence of mus-cle hypertonia is the activation of anaerobic glycolysis, accompanied by the accumulation of lactic acid, which

d e n t i s t r y

http://dx.doi.org/10.35630/2199-885X/2021/11/2/31

https://orcid.org/0000-0002-5546-8570

https://orcid.org/0000-0001-8378-0505

https://orcid.org/0000-0002-4648-7569

https://orcid.org/0000-0003-1742-9455

https://orcid.org/0000-0002-1905-7723

https://orcid.org/0000-0002-0643-6863

http://dx.doi.org/10.35630/2199-885X/2021/11/2/31


R E Q U I R E M E N T S A N D I N D I C A T I O N SIn the available literature, we did not find the

requirements for pneumatic trainers — rotary dilators, so we tried to formulate them based on their structure and function under normal conditions.

From our point of view, a pneumatic trainer should:

— provide therapeutic and prophylactic effects;— transfer action to the chewing muscles;— have a range of motion close to that of a healthy

person;— have a lightweight and durable design, adaptable

to the anatomical parameters of the oral cavity;— be bioinert and safe for the patient;— carry out the expansion of the mouth by the

amount of 40–50 mm;— be elastic and durable, to maintain high intracavi-

tary pressure;— be able to cyclically dosed air injection to the

required values and its release as needed, depend-ing on the nature of the disease and the impact on it;

— provide adequate access to the dentition for therapeutic and prophylactic treatment by a doc-tor;

— possess simplicity and availability of activation for a doctor and a patient;

— exclude the possibility of slipping from the denti-tion;

— exclude the possibility of aspiration during ma-nipulations;

— to be easily disinfected, cleaned and sterilized;— have the possibility of individual anatomical

parameterization;— do not affect the observance of oral hygiene;— be easily to withdraw in case of inflammation of

the mucous membrane and injury to the gingival papillae;

— be mobile and independent of power sources and affordable for the mass consumer.

This device should only be used and prescribed by compliant patients making regular visits to a dentist.

The use of a pneumatic expander is indicated for the following diseases classified according to the ICD-10 code.

Many patients with TMJ do not go to the doctor, however, they note the presence of one or more of the following symptoms, which are indication for the use of a pneumatic expander.

— Muscle pain;— Night grinding of teeth;— Pathological abrasion of teeth, chips of ortho-

pedic and therapeutic structures;

— Fatigue of the chewing and facial muscles;— Pathological bite;— Headaches, migraines, tinnitus and ringing in

the ears caused by overexertion of the chewing muscles;

— Abnormal fixed postural reflexes;— Restriction of mouth opening;— Pain in the joint while eating, talking;— Change of face configuration due to hyper; masti-

catory muscle atrophy.

C O N T R A I N D I C A T I O N SThere are also absolute and relative contraindica-

tions for a gnatic device:Absolute:

— Intolerance to the components of the pneumatic expander-retractor;

— Increased gag reflex;Relative:— Low level of compliance between parents and

child;— Acute respiratory diseases;— Unstable psycho-emotional behavior of the child;— Persistent contracture of the temporomandibular

joint, with limited mouth opening less than 10 mm;

1. Dysfunctional conditions of the TMJ:

K07.6 – diseases of the temporomandibular joint (neuromuscular dysfunctional syndrome, occlusive-articulation syndrome, pain dysfunc-tion syndrome)K07.62 – Habitual dislocation and subluxation of mandibleK07.64 – Temporomandibular joint stiffnessS03.0 – dislocation of the intra-articular meniscusS03.4 – Temporomandibular joint ligament sprain

2. Arthritis:

M00.VX – pyogenic arthritis of the TMJM12.5X – traumatic arthropathy (acute traumatic arthritis)M05.VX – rheumatoid, rheumatic, infectious-allergic arthritis

3. Arthrosis: M19.0X – primary arthrosis of other joints (TMJ)M24.6VX – joint ankylosis

4. Muscle damage:

F45.8 – Other somatoform disorders (bruxism, teeth grinding)M60.0 – Infectious myositis of the masticatory musclesM62.5 – Muscle wasting and wasting, not elsewhere classifiedM24.8.0 – muscle hypertonicity

5. Diseases of the nervous system:

G43 – MigraineG43.2 – Tension type headacheK14.6 – Glossodyny

d e n t i s t r y


— Period of exacerbation of periodontal and tempo-romandibular joint diseases;

— Cheilitis and erosive and ulcerative diseases of the oral mucosa in the acute stage.

C O N C L U S I O NThus, clinical effects of the use of a pneumatic

expander are achieved by redistributing the occlusal load on the joint, which results in gradually normal-izing the function of masticatory muscles. This may be seen as a solution to myology problems in oral and maxillofacial region.

The application of a multifunctional gnathic device can be complemented with exercises and relax-ing techniques to improve the therapeutic effect on the masticatory muscles.

R E F E R E N C E S1. Bergmann A., Edelhoff D., Schubert O.,

Erdelt K.J. Effect of treatment with a full-occlusion biofeedback splint on sleep bruxism and TMD pain: a randomized controlled clinical trial // Clin Oral Investig. – 2020. – Vol.24 – No. 11 – P. 4005–4018. doi: 10.1007 / s00784-020-03270-z.

2. Connaghan K.P., Moore C.A. Indirect estimates of jaw muscle tension in children with suspected hypertonia, children with suspected hypotonia, and matched controls // J Speech Lang Hear Res. – 2013 – Vol. 56. – no. 1. – P.123–136. doi: 10.1044/1092-4388(2012/11-0161).

3. Avanisyan V., Al-Harazi G., Kondratyeva T., Harutyunyan Yu. Morphology of facial skeleton in children with undifferentiated connec-tive tissue dysplasia. Archiv EuroMedica. 2020. Vol. 10; 3: 130–141. https://dx.doi.org/10.35630/2199-885X/2020/10/3.32

4. Dmitrienko S.V. Analytical approach within cephalometric studies assessment in people with various somatotypes. Archiv EuroMedica. 2019. Vol. 9; 3: 103–111. https://doi.org/10.35630/2199-885X/2019/9/3.29

5. Domenyuk D.A. Contemporary methodological approaches to diagnosing bone tissue disturbances in children with type 1 diabetes. Archiv Euro-Medica, 2018; 8(2): 71–81. DOI:10.35630/2199-885x/2018/8/2/71

6. Harutyunyan Yu. Undifferentiated connective tissue dysplasia as a key factor in pathogenesis of maxil-lofacial disorders in children and adolescents. Archiv EuroMedica. 2020. Vol. 10; 2: 83–94. https://dx.doi.org/10.35630/2199-885X/2020/10/2.24

7. Ivanyuta O.P., Al-Harasi G., Kuleshov D.A. Modification of the dental arch shape using graphic reproduction method and its clinical effectiveness in patients with occlusion anomalies. Archiv Euro-Medica. 2020. Vol. 10; 4: 181–190. https://dx.doi.org/10.35630/2199-885X/2020/10/4.42

8. Shkarin V.V., Kochkonyan T.S., Ghamdan Al.H. Occlusal plane orientation in patients with dentofacial anomalies based on morphometric cranio-facial measurements. Archiv EuroMedica. 2021. Vol. 11; 1: 116–121. https://doi.org/10.35630/2199-885X/2021/11/1.26

9. Poroyskiy S.V., Makedonova Yu.A., Kurkina O.N., Shkarin V.V. Gingivectomy as a method of preparation for orthopedic treatment in patients with bottom dental crown of abutment teeth.// Archiv Euromedica. – 2020. – No. 2 (10). – p. 124–126

10. Santana U., Mora M.J. Electromyographic analysis of the masticatory muscles of patients after com-plete rehabilitation of occlusion with protection by non-working side contacts // J Oral Rehabil.-1995 – Vol. 22. – No.1. – P. 57–66. doi: 10.1111 / j.1365-2842.1995.tb00211.x. PMID: 7897560.

11. Soikher M.I., Orlova O.R., Mingazova L.R., et al. Comparative prospective study of the efficacy and safety of the Russian preparation of botulinum toxin type A relatox® for the purpose of correcting hypertonicity of the masticatory muscles in patients with myofascial pain syndrome // Russian dental jour-nal. – 2019. – Vol. 23. – No. 3–4. – P. 180–184. DOI: 10.18821 / 1728-2802-2019-23-3-4-180-184

12. Vorobev A.A., Makedonova Yu. A., Aleksan-drov A. V., Sosulya E. Yu. Gnatic device / Patent No.2020116273/14 (026641)

13. Vorobyev A. A., Chigrova N. A., Pylaeva I. O., Barinova E. A. Cosmetological anatomy of the face. 2nd edition, expanded and revise // Publishing house "Elbi" St. Petersburg. 2019, 340 p.

14. Fadeev R.A., Ovsyannikov K.A. Etiology and pathogenesis of diseases of the temporomandibular joint and masticatory muscles // Bulletin of the Novgorod State University. Yaroslav the Wise. – 2020. – No. 4 (120) – P. 50–59. DOI: 10.34680 / 2076-8052.2020.4 (120) .50-59

d e n t i s t r y